Composition for predicting prognosis of thyroid cancer

Abstract

The present invention relates to a combination of markers that enables the prognosis of thyroid cancer to be predicted with high accuracy before surgical treatment is performed on a thyroid nodule, thereby allowing a determination as to whether to perform the surgery to be made in advance. Since the prognosis of thyroid cancer can thus be predicted at an earlier stage and with greater precision, the combination of markers is expected to minimize unnecessary surgery on a thyroid nodule.

Description

Composition for predicting the prognosis of thyroid cancer

[0001] The present invention relates to a combination of biomarkers for predicting the prognosis of thyroid cancer.

[0002] Thyroid cancer refers to cancer that develops in the thyroid gland and includes papillary carcinoma, follicular carcinoma, Hurthle cell neoplasm, anaplastic carcinoma, and medullary thyroid carcinoma. According to the 2019 National Cancer Registry Statistics, thyroid cancer was reported to be the most common type of new cancer patient. It is a cancer that occurs more frequently in women than in men, and over the past five years, it has been identified as the number one cancer with the highest incidence rate among young patients (ages 15 to 34).

[0003] Although radiation exposure and sex hormones are known causes of thyroid cancer, the exact cause has not yet been identified, and it is known that, as with other cancers, various genes and genetic mutations are involved. Although the incidence of thyroid cancer is higher than that of other cancers, the survival rate is relatively high if surgery is performed early; therefore, early detection is important, and various attempts are being made to detect thyroid cancer at an early stage.

[0004] The diagnosis of thyroid cancer requires a combination of clinical screening, imaging tests such as ultrasound, and cytopathological examinations such as fine needle aspiration (FNA). FNA is the most important test for diagnosing thyroid cancer because it can distinguish between benign and malignant nodules with high accuracy.

[0005] However, up to 30% of Fine Needle Aspiration (FNA) results are indeterminate nodules whose diagnosis is uncertain based solely on cytopathological examination, making clinical treatment decisions difficult. Furthermore, there is a limitation in that 15–35% of patients who undergo surgery for indeterminate nodules are diagnosed as malignant afterward, resulting in the remaining 65–85% receiving unnecessary surgery. Therefore, molecular marker testing is essential to accurately distinguish thyroid cancer from benign tumors. Recently, research aimed at discovering molecular markers capable of precisely diagnosing thyroid cancer has been actively conducted. Currently available panel-based tests improve the diagnostic accuracy of indeterminate nodules by collecting mutation information (SNV / INDEL, CNV, Gene Fusion, Overexpression) from small amounts of DNA / RNA obtained from thyroid nodules; however, research on markers that can determine whether surgical treatment is necessary is lacking.

[0006] Accordingly, the inventors have arrived at the present invention by discovering a combination of markers that can help determine whether to perform surgery in advance with high accuracy by predicting the prognosis (risk of malignancy) of thyroid cancer before proceeding with surgical treatment for thyroid nodules.

[0007] The present invention aims to provide an optimal combination of prognostic biomarkers suitable for distinguishing patients predicted to have a high or low risk of malignancy, in order to assist in making decisions regarding surgical treatment for thyroid nodules.

[0008] As a result of diligent efforts to achieve the above objective, the inventors have discovered a combination of prognostic biomarkers capable of determining whether to perform surgical treatment for thyroid nodules by searching for genes that have a clinical association with thyroid cancer.

[0009] One objective of the present invention is to provide a composition for predicting the prognosis of thyroid cancer.

[0010] Another objective of the present invention is to provide a kit for predicting the prognosis of thyroid cancer.

[0011] Another objective of the present invention is to provide a method for providing information for predicting the prognosis of thyroid cancer.

[0012] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below.

[0013] Various embodiments of the present invention are described with reference to the drawings. In the following description, for a complete understanding of the present invention, various specific details, such as specific forms, compositions, and processes, are described. However, specific embodiments may be practiced without one or more of these specific details, or in combination with other known methods and forms. In other examples, known processes and manufacturing techniques are not described as specific details so as not to make the present invention unnecessary or obscure. Reference throughout this specification to one embodiment implies that the particular features, forms, compositions, or characteristics described in association with the embodiment are included in one or more embodiments of the present invention. Accordingly, the circumstances of the embodiments expressed at various locations throughout this specification do not necessarily represent the same embodiment of the present invention. Additionally, particular features, forms, compositions, or characteristics may be combined in any suitable way in one or more embodiments. Unless otherwise defined in the specification, all scientific and technical terms used in this specification have the same meaning as commonly understood by those skilled in the art to which the present invention pertains.

[0014]

[0015] According to one embodiment of the present invention, the invention relates to a composition for predicting the prognosis of cancer.

[0016] In the present invention, the composition may include a preparation for detecting one or more mutations on genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

[0017] In the present invention, the information on the "BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT genes" is registered with the NCBI (National Center for Biotechnology Information), and the disclosed Gene ID information of the said genes of the present invention is as follows, and may include all of them without limitation within the scope where it is obvious to a person skilled in the art that they produce the intended effects of the present invention:

[0018] BRAF (Gene ID: 673), KRAS (Gene ID: 3845), HRAS (Gene ID: 3265), NRAS (Gene ID: 4893), PIK3CA (Gene ID: 5290), RET (Gene ID: 5979), TP53 (Gene ID: 7157) and TERT (Gene ID: 7015)

[0019] In the present invention, the composition is intended to be applied to a biological sample isolated from a target individual, and the biological sample may include, but is not limited to, solid tissue samples, tissue culture media, liquid tissue samples, cells, or cell fragments. In addition, as non-limiting examples of biological samples, whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, and joint aspirate It may include one or more selected from the group consisting of aspirate, organ secretions, cell, cell extract, and cerebrospinal fluid, but is not limited thereto.

[0020] In the present invention, the term "intended individual" refers to an individual that has developed cancer or has a high probability of developing cancer, and may be a mammal including humans, and may be selected from a group consisting of, for example, humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, dogs, cats, cattle, horses, pigs, sheep, and goats, and specifically, for the purposes of this project, may be a patient with thyroid cancer, but is not limited thereto.

[0021] When a mutation of the gene according to the present invention is detected in a biological sample isolated from the target individual in the present invention, the prognosis of the disease can be predicted very quickly and simply, and furthermore, it can be determined whether to proceed with surgical treatment.

[0022] In the present invention, the "nodule" is classified according to its cause and histological characteristics into hyperplasia nodules, colloid nodules, inflammatory nodules, thyroid cysts, and neoplastic nodules including thyroid cancer. Although distinguishing between thyroid cancer and benign nodules is of utmost importance in the diagnosis of thyroid nodules, there is a problem in that accurate diagnosis is difficult when the thyroid nodule falls into the indeterminate nodule category in the case of fine-needle aspiration cytology, which is currently widely performed for the diagnosis of thyroid cancer. For the purposes of the present invention, there is an effect of accurately predicting whether a thyroid nodule requires surgery.

[0023] In the present invention, the term "cancer" refers to or indicates a physiological condition characterized by uncontrolled cell growth in mammals. In the present invention, the cancer may be thyroid cancer, parathyroid cancer, pancreatic cancer, stomach cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, pro-anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma, and specifically may be thyroid cancer, but the progression of cancer, such as differentiation and / or proliferation of the tumor, in the present invention This is not limited to types of cancer that depend on the cancer cells and / or cancer stem cells described.

[0024] In the present invention, the term "thyroid cancer" refers to cancer that develops in the thyroid gland and is the most common endocrine tumor. Such thyroid cancers include papillary carcinomas and follicular carcinomas, which have a good prognosis, as well as poorly differentiated and undifferentiated carcinomas, which have a poor prognosis; in addition, there are medullary carcinomas, lymphomas, etc.

[0025] In the present invention, the term "prognosis" refers to the act of predicting the risk of malignancy of a disease in advance, and furthermore, it may involve predicting the ease of treatment (necessity of surgical treatment) by anticipating the future course or outcome of cancer. It may involve distinguishing cancer from other diseases, for example, thyroid diseases, particularly thyroiditis (including both acute and chronic forms), or benign thyroid tumors (such as thyroid nodules or lumps that are not neoplastic nodules), or distinguishing carcinomas, particularly distinguishing thyroid cancer from other carcinomas. The above prognosis or prognosis prediction may be interpreted as meaning any act of predicting the course of a disease prior to treatment by comprehensively considering the patient's condition, as the course of the disease may vary depending on the patient's physiological or environmental state. For the purposes of the present invention, the above prognosis prediction may involve predicting the possibility of progression or risk of malignancy prior to surgical treatment of cancer, or it may involve predicting whether surgical treatment of cancer is necessary. Specifically, a good prognosis means that the nodule has a low risk of malignancy and is likely benign, indicating a low need for surgery. A poor prognosis means that the nodule has a high risk of malignancy and is likely malignant, indicating a high need for surgery.

[0026] In the present invention, the term “treatment” refers to any act that provides clinically beneficial effects on cancer or a disease in its pre-disease stage in a target individual. This is not limited to the mere alleviation of symptoms but may include all medical, surgical, pharmacological, or supportive measures that prevent or suppress the occurrence, progression, and recurrence of the disease, and prevent or improve the exacerbation of an already developed disease. Specifically, the “treatment” may include prophylactic treatment to lower or delay the likelihood of cancer development in individuals at high risk of developing cancer; surgical treatment to directly remove cancer or inhibit its progression by resecting tumors or nodules; pharmacological treatment to inhibit the growth or differentiation of cancer cells or induce apoptosis by administering drugs such as anticancer agents, targeted therapies, immunotherapies, or hormones; treatment utilizing physical energy such as radiation, radiofrequency, or lasers; and supportive treatment to improve prognosis and prevent recurrence by administering it before or after surgery or in conjunction with other treatments. In addition, the present invention may include prognosis-based treatment that predicts the prognosis through the detection of gene mutations, determines the risk of malignancy and the necessity of surgery based on the results, and selects and applies an appropriate treatment strategy.

[0027] In the present invention, the term "tumor" refers to all neoplastic cell growth and proliferation, all precancerous states, and cancerous cells and tissues, regardless of whether they are malignant or benign. In the present invention, tumors may be pathologically classified into benign tumors, borderline tumors, well-differentiated tumors, and de-differentiated tumors, but are not limited thereto.

[0028] In the present invention, the term "benign thyroid tumor" refers to a benign tumor among various types of thyroid nodules in which a part of the tissue enlarges due to the overproliferation of thyroid cells to form a lump, excluding neoplastic nodules containing cancer.

[0029] In the present invention, the agent for detecting the mutation may include one or more selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to a protein encoded by a gene containing a mutation site on the gene to be detected, but is not limited thereto.

[0030] In the present invention, the term "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, an antibody means an antibody that specifically binds to the protein. The antibodies of the present invention include polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. The antibodies can be easily manufactured using techniques widely known in the art. For example, polyclonal antibodies can be produced by a method widely known in the art that includes the process of injecting the antigen of the protein into an animal and collecting blood from the animal to obtain serum containing antibodies. Such polyclonal antibodies can be produced from any animal, such as goats, rabbits, sheep, monkeys, horses, pigs, cattle, dogs, etc. In addition, monoclonal antibodies may be prepared using the hybridoma method (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), which is widely known in the industry, or phage antibody library technology (see Clackson et al, Nature, 352:624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above methods may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. Furthermore, the antibodies of the present invention comprise not only a complete form having two full-length light chains and two full-length heavy chains, but also functional fragments of the antibody molecule. A functional fragment of an antibody molecule refers to a fragment that possesses at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

[0031] In the present invention, the "oligopeptide" is a peptide composed of 2 to 20 amino acids and may include dipeptides, tripeptides, tetrapeptides, and pentapeptides, but is not limited thereto.

[0032] In the present invention, the "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized polymer similar to DNA or RNA, which was first introduced in 1991 by Professors Nielsen, Egholm, Berg, and Buchardt of the University of Copenhagen, Denmark. While DNA has a phosphate-ribose sugar backbone, PNA has a repeating N-(2-aminoethyl)-glycine backbone connected by peptide bonds, which significantly increases its binding affinity and stability to DNA or RNA, and is therefore used in molecular biology, diagnostic analysis, and antisense therapy. PNA is disclosed in detail in the literature [Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide". Science 254 (5037): 1497-1500].

[0033] In the present invention, the "aptamer" is an oligonucleotide or peptide molecule, and general information regarding aptamers is disclosed in detail in the literature [Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727(1998)].

[0034] In the present invention, the agent for detecting the mutation may include one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to a mutation site on the gene to be detected, but is not limited thereto.

[0035] In the present invention, the "primer" is a fragment that recognizes a target gene sequence and includes a forward and reverse primer pair, but preferably is a primer pair that provides analysis results having specificity and sensitivity. High specificity can be conferred when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that it amplifies only the target gene sequence containing the complementary primer binding site and does not induce non-specific amplification.

[0036] In the present invention, the term "probe" refers to a substance capable of specifically binding to a target substance to be detected within a sample, and means a substance capable of specifically confirming the presence of the target substance within the sample through said binding. The type of probe is not limited to substances commonly used in the industry, but preferably may be PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA, or DNA, and most preferably PNA. More specifically, the probe may be a biomaterial derived from an organism or similar, or manufactured in vitro, and may be, for example, enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA, and RNA; DNA may include cDNA, genomic DNA, and oligonucleotides; RNA may include genomic RNA, mRNA, and oligonucleotides; and examples of proteins may include antibodies, antigens, enzymes, peptides, etc.

[0037] In the present invention, "LNA (Locked nucleic acids)" refers to nucleic acid analogs containing a 2'-O, 4'-C methylene bridge [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502]. LNA nucleosides contain common nucleic acid bases of DNA and RNA and can form base pairs according to the Watson-Crick base pairing rule. However, due to the 'locking' of the molecule caused by the methylene bridge, LNAs are unable to form an ideal shape in Watson-Crick bonding. When LNAs are included in DNA or RNA oligonucleotides, LNAs can pair more quickly with complementary nucleotide chains, thereby increasing the stability of the double helix.

[0038] In the present invention, "antisense" refers to an oligomer having a backbone between nucleotide base sequences and subunits, wherein the antisense oligomer hybridizes with a target sequence within RNA by Watson-Crick base pairing, thereby allowing the formation of a mRNA and RNA:oligomer heterodimer within the target sequence. The oligomer may have exact sequence complementarity or approximate complementarity with respect to the target sequence.

[0039] In the present invention, the "mutation" is a permanent change in the base sequence of an existing gene and may be one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, inversion, and gene fusion, and specifically may be a single nucleotide variation (SNV), an insertion / deletion variation (Indel), or a gene fusion, but is not limited thereto.

[0040] In one embodiment of the present invention, the BRAF mutation is characterized as being a variant gene (p.V600E) in which valine (V) at position 600 of the amino acid sequence of SEQ ID NO. 1 is substituted with glutamic acid (E).

[0041] In another embodiment of the present invention, the KRAS mutation is characterized as being a variant gene (p.G12) in which glycine (G) at position 12 of the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid.

[0042] In another embodiment of the present invention, the HRAS mutation is characterized as being a variant gene (p.G13) in which glycine (G) at position 13 of the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid.

[0043] In another embodiment of the present invention, the NRAS mutation is characterized as being a variant gene (p.Q61) in which glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid.

[0044] In another embodiment of the present invention, the PIK3CA mutation is characterized by being one or more selected from the group consisting of: a variant gene (p.E545K) in which glutamic acid (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K); and a variant gene (p.H1047R) in which histidine (H) at position 1047 in the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R).

[0045] In another embodiment of the present invention, the RET mutation is characterized by being one or more selected from the group consisting of: a variant gene (p. C634) in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; and a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T).

[0046] In another embodiment of the present invention, the TP53 mutation is characterized by being one or more selected from the group consisting of: a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); and a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid.

[0047] In another embodiment of the present invention, the TERT mutation is characterized by being one or more selected from the group consisting of: a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,135 (based on hg38)).

[0048] The composition of the present invention may further include a preparation for detecting mutations on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

[0049] In one embodiment of the present invention, the RET mutation may be a fusion gene in which the RET gene appears in a fused form with another gene, and the other gene may be the CCDC6 gene, but is not limited thereto.

[0050] In another embodiment of the present invention, the BRAF mutation may be a fusion gene in which the BRAF gene appears in a fused form with another gene, and the other gene may be the AKAP9 gene, but is not limited thereto.

[0051] In another embodiment of the present invention, the NTRK1 mutation may be a fusion gene in which the NTRK1 gene appears in a fused form with another gene, and the other gene may be the TPM3 gene, but is not limited thereto.

[0052] Since the information on BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, TERT, and NTRK1 according to the present invention, or the genes encoding them, is known, a person skilled in the art can easily design primers, probes, or antisense nucleotides that specifically bind to said mutant proteins or mutant genes based on this.

[0053] According to another embodiment of the present invention, the present invention relates to a kit for predicting the prognosis of cancer comprising the composition for predicting the prognosis of the present invention.

[0054] In the present invention, the "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. It includes not only direct labeling of a detectable substance related to a probe or antibody through reaction with a substrate, but also indirect labeling in which a label that develops color through reactivity with another directly labeled reagent is conjugated. It may include a color-developing substrate solution, washing solution, and other solutions that react with the label for color development, and may be manufactured to include the reagent components used. In the present invention, the kit may be a kit containing essential elements necessary for performing RT-PCR, and in addition to each primer pair specific to the marker gene, it may include test tubes, reaction buffer, deoxyribonucleotides (dNTPs), Taq polymerase, reverse transcriptase, DNase, RNase inhibitor, sterile water, etc. Furthermore, the kit may be a kit for detecting genes for cancer diagnosis that includes essential elements necessary for performing DNA chip analysis. A DNA chip kit comprises a substrate to which cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may comprise cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, provided that it is known in the art.

[0055] In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0056] The kit of the present invention may further include one or more other component compositions, solutions, or devices suitable for the analysis method. For example, the kit of the present invention may further include essential elements necessary to perform a reverse transcription polymerase chain reaction. The reverse transcription polymerase chain reaction kit includes a primer pair specific to a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also include a primer specific to the nucleic acid sequence of a control gene. Furthermore, the reverse transcription polymerase chain reaction kit may include a test tube or other suitable container, a reaction buffer (with varying pH and magnesium concentration), deoxyribonucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC-water, sterile water, etc.

[0057] In addition, the cancer prognosis prediction kit of the present invention may include essential elements necessary for performing DNA chip operations. The DNA chip kit may include a substrate to which cDNA or oligonucleotides corresponding to a gene or a fragment thereof are attached, and reagents, preparations, enzymes, etc., for producing fluorescently labeled probes. Additionally, the substrate may include cDNA or oligonucleotides corresponding to a control gene or a fragment thereof.

[0058] In addition, the cancer prognosis prediction kit of the present invention may include essential elements necessary for performing ELISA. The ELISA kit includes an antibody specific to the protein. The antibody is an antibody that has high specificity and affinity for the marker protein and has little cross-reactivity with other proteins, and is a monoclonal antibody, a polyclonal antibody, or a recombinant antibody. Additionally, the ELISA kit may include an antibody specific to a control protein. Furthermore, the ELISA kit may include reagents capable of detecting the bound antibody, such as a labeled secondary antibody, chromophores, an enzyme (e.g., conjugated with the antibody) and its substrate, or other substances capable of binding to the antibody.

[0059] In the present invention, the immobilizer for the antigen-antibody binding reaction may be a nitrocellulose membrane, a PVDF membrane, a well plate synthesized from polyvinyl resin or polystyrene resin, a glass slide glass, etc., but is not limited thereto.

[0060] In addition, in the present invention, the label of the secondary antibody is preferably a conventional chromogenic agent that undergoes a color reaction, and may be labeled with fluorescent substances (fluorescein) and dyes such as HRP (horseradish peroxidase), alkaline phosphatase, colloid gold, FITC (poly L-lysine-fluorescein isothiocyanate), RITC (rhodamine-B-isothiocyanate), and Europium chelate, but is not limited thereto.

[0061] In addition, in the present invention, it is preferable to use a chromogenic substrate to induce color development depending on the label that performs the color reaction, and TMB (3,3',5,5'-tetramethylbezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD (o-phenylenediamine), etc. may be used. At this time, it is more preferable that the chromogenic substrate be provided in a state dissolved in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is degraded by HRP used as a label for the secondary antibody conjugate to produce a chromogenic precipitate, and the presence or absence of the marker proteins is detected by visually confirming the degree of precipitation of this chromogenic precipitate.

[0062] In the present invention, the washing solution preferably comprises a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20 is more preferably used. After the antigen-antibody binding reaction, a secondary antibody is reacted with the antigen-antibody conjugate, and then an appropriate amount of the washing solution is added to the immobilizer to wash 3 to 6 times. A sulfuric acid solution (H2SO4) may preferably be used as the reaction stopping solution.

[0063] According to another embodiment of the present invention, the invention relates to a method for providing information for predicting the prognosis of cancer.

[0064] The method of the present invention may include the step of detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT in a biological sample isolated from a target individual.

[0065] The method of the present invention may further include the step of detecting a mutation on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

[0066] In the present invention, the object of the above invention is an object that has developed cancer or is highly likely to develop cancer, and may be a mammal including humans, and may be selected from the group consisting of, for example, humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, dogs, cats, cattle, horses, pigs, sheep, and goats, and preferably may be a human, but is not limited thereto.

[0067] In the present invention, the biological sample refers to any substance, biological body fluid, tissue, or cell obtained from or derived from an individual, including whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, and bronchial aspirate. It may be one or more selected from the group consisting of synovial fluid, joint aspirate, organ secretions, cell, cell extract and cerebrospinal fluid, but is not limited thereto.

[0068] In the present invention, the agent for detecting mutations on the gene may include one or more selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to a protein encoded by a gene containing a mutation site on the gene.

[0069] In the present invention, the detection of the mutant protein may be performed by protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Surface Enhanced Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion, Ouchteroni immunodiffusion, Rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry (LC-MS), LC-MS / MS (liquid chromatography-mass spectrometry / mass spectrometry), Western blotting, or ELISA (enzyme-linked immunosorbent assay).

[0070] In the present invention, for the multiple reaction monitoring method, the internal standard substance may be a synthetic peptide in which a specific amino acid constituting the target peptide is substituted with an isotope, or E. coli beta-galactosidase.

[0071] In the present invention, the agent for detecting mutations on the gene may include one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to mutation sites on the gene.

[0072] In the present invention, the detection of the gene mutation may be performed by reverse transcription polymerase chain reaction (RT-PCR), competitive reverse transcription polymerase chain reaction (Competitive RT-PCR), real-time reverse transcription polymerase chain reaction (Real-time RT-PCR), RNase protection assay (RPA), Northern blotting, or a DNA chip.

[0073] In one embodiment of the present invention, the method comprises a mutant gene detected in a biological sample of the target individual, wherein the mutant gene is a variant gene (p.V600E) in which valine (V) at position 600 of the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E); a variant gene (p.G12) in which glycine (G) at position 12 of the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid; a variant gene (p.G13) in which glycine (G) at position 13 of the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid; a variant gene (p.Q61) in which glutamine (Q) at position 61 of the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid; and a variant gene (p.E545K) in which glutamate (E) at position 545 of the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K). A variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); a variant gene (p. C634) in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; If one or more types are selected from the group consisting of a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,135 (based on hg38)); the prognosis of the cancer can be predicted to be poor.

[0074] In another embodiment of the present invention, the method can predict that the prognosis of cancer will be poor if the mutant gene detected in the biological sample of the target individual is one or more selected from the group consisting of: a fusion gene in which the RET gene is fused with the CCDC6 gene; a fusion gene in which the BRAF gene is fused with the AKAP9 gene; and a fusion gene in which the NTRK1 gene is fused with the TPM3 gene.

[0075] In the above method of the present invention, descriptions regarding mutation information of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, TERT, and NTRK1, cancer, antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), aptamers, etc., and descriptions regarding primers, probes, etc. are duplicated from what has been previously described, so in order to avoid excessive complexity of the specification, detailed descriptions thereof are omitted below.

[0076] In a first aspect of the present invention, a composition for predicting the prognosis of cancer is provided, comprising a preparation for detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

[0077] In the first embodiment above, the second embodiment provides a composition in which the preparation comprises one or more selected from the group consisting of: a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

[0078] In the first or second embodiment above, the third embodiment provides a composition in which the mutation is one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, and inversion.

[0079] In any one of the first to third embodiments above, the fourth embodiment provides a composition characterized in that the BRAF mutation is a variant gene (p.V600E) in which valine (V) at position 600 in the amino acid sequence of SEQ ID NO. 1 is substituted with glutamic acid (E); in any one of the first to fourth embodiments above, the fifth embodiment provides a composition characterized in that the KRAS mutation is a variant gene (p.G12) in which glycine (G) at position 12 in the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid; and in any one of the first to fifth embodiments above, the sixth embodiment provides a composition characterized in that the HRAS mutation is a variant gene (p.G13) in which glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid. In any one of the embodiments 1 to 6, the seventh embodiment is characterized in that, in claim 1, the NRAS mutation is a variant gene (p.Q61) in which glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid; and in any one of the embodiments 1 to 7, the eighth embodiment is characterized in that the PIK3CA mutation is a variant gene (p.E545K) in which glutamic acid (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K); The present invention provides a composition characterized by comprising one or more selected from the group consisting of: a variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); wherein, in any one of the first to eighth embodiments, the ninth embodiment comprises a variant gene (p. in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid.The present invention provides a composition characterized by comprising one or more selected from the group consisting of C634); and a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); wherein, in any one of the first to ninth embodiments, the tenth embodiment comprises a TP53 mutation in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H) (p. R175H); The present invention provides a composition characterized by being one or more selected from the group consisting of: a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; and in any one of the first to tenth embodiments, the eleventh embodiment is characterized in that the TERT mutation is one or more selected from the group consisting of: a promoter variant gene (chr5: 1,295,113 (based on hg38)) in which cytosine (C), which is the complementary base sequence of position 127 of SEQ ID NO. 8, is substituted with thymine (T); and a promoter variant gene (chr5: 1,295,135 (based on hg38)) in which cytosine (C), which is the complementary base sequence of position 149 of SEQ ID NO. 8, is substituted with thymine (T).

[0080] In any one of the first to eleven embodiments above, the twelfth embodiment provides a composition in which the composition further comprises a preparation for detecting mutations on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

[0081] In any one of the first to twelfth embodiments above, the twelfth embodiment provides a composition in which the RET mutation appears in a form in which the RET gene is fused with CCDC6, and in any one of the first to twelfth embodiments above, the twelfth embodiment provides a composition in which the BRAF mutation appears in a form in which the BRAF gene is fused with AKAP9, and in any one of the first to twelfth embodiments above, the twelfth embodiment provides a composition in which the NTRK1 mutation appears in a form in which the NTRK1 gene is fused with TPM3.

[0082] In any one of the first to fifteen embodiments above, the sixth embodiment is that the cancer is thyroid cancer, parathyroid cancer, pancreatic cancer, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, prostatic cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma. A composition is provided, at least one selected from the group.

[0083] The 17th embodiment provides a kit for predicting the prognosis of cancer, comprising a composition of any one of the 1st to 16th embodiments.

[0084] In the 18th aspect of the present invention, a method for providing information regarding the prognosis prediction of cancer is provided, comprising the step of detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT in a biological sample isolated from a target individual.

[0085] In the above 18th embodiment, the 19th embodiment provides a method comprising the step of detecting a mutation on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

[0086] In the above 18th or 19th embodiment, the 20th embodiment comprises: a variant gene (p.V600E) in which valine (V) at position 600 in the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E); a variant gene (p.G12) in which glycine (G) at position 12 in the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid; a variant gene (p.G13) in which glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid; a variant gene (p.Q61) in which glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid; and a variant gene (p.E545K) in which glutamate (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K). A variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); a variant gene (p. C634) in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; If one or more are selected from the group consisting of: a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (hg38 기준)); and a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,135 (hg38 기준));

[0087] The above-mentioned entity provides a method characterized by predicting that the prognosis of cancer will be poor.

[0088] In any one of the 18th to 20th embodiments, the 21st embodiment provides a method characterized in that, when the mutant gene is one or more selected from the group consisting of a fusion gene in which the RET gene is fused with the CCDC6 gene; a fusion gene in which the BRAF gene is fused with the AKAP9 gene; and a fusion gene in which the NTRK1 gene is fused with the TPM3 gene; the individual is predicted to have a poor prognosis for cancer.

[0089] In any one of the 18th to 21st embodiments, the 22nd embodiment is that the cancer is thyroid cancer, parathyroid cancer, pancreatic cancer, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, prostatic cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma. Provides a method, at least one selected from the military.

[0090] In the 23rd aspect of the present invention, the use of a preparation for cancer prognosis prediction is provided for detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

[0091] In the above 23rd embodiment, the 24th embodiment provides a use for predicting cancer prognosis, wherein the preparation comprises one or more selected from the group consisting of a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

[0092] In the above 23rd or 24th embodiment, the 25th embodiment provides a cancer prognosis prediction use in which the mutation is one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, and inversion; in any one of the above 23rd to 25th embodiments, the 26th embodiment provides a cancer prognosis prediction use in which the BRAF mutation is a variant gene (p.V600E) in which valine (V) at position 600 of the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E); and in any one of the above 23rd to 26th embodiments, the 27th embodiment provides a KRAS mutation in which the amino acid of SEQ ID NO. 2 The present invention provides a cancer prognosis prediction application in which the glycine (G) at position 12 in the sequence is substituted with another amino acid, wherein in any one of the 23rd to 27th embodiments, the 28th embodiment provides a cancer prognosis prediction application in which the HRAS mutation is a cancer prognosis prediction application in which the glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid, wherein in any one of the 23rd to 28th embodiments, the 29th embodiment provides a cancer prognosis prediction application in which the NRAS mutation is a cancer prognosis prediction application in which the glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid, wherein in any one of the 23rd to 29th embodiments, the 30th embodiment provides a cancer prognosis prediction application in which the PIK3CA mutation is SEQ ID NO. A variant gene (p. in which glutamic acid (E) at position 545 of the amino acid sequence of 5 is replaced with lysine (K).The invention provides a use for predicting cancer prognosis, comprising one or more selected from the group consisting of E545K); and a variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); wherein, in any one of the 23rd to 30th embodiments, the 31st embodiment comprises a RET mutation in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid (p. C634); The invention provides a use for predicting cancer prognosis, comprising one or more selected from the group consisting of: a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); wherein, in any one of the 23rd to 31st embodiments, the 32nd embodiment comprises a TP53 mutation in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); The present invention provides a cancer prognosis prediction use selected from the group consisting of: a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; and in any one of the 23rd to 32nd embodiments, the 33rd embodiment provides a cancer prognosis prediction use selected from the group consisting of: a promoter variant gene (chr5: 1,295,113 (hg38 기준)) in which cytosine (C) at position 127 of the complementary nucleotide sequence of SEQ ID NO. 8 is substituted with thymine (T); and a promoter variant gene (chr5: 1,295,135 (hg38 기준)) in which cytosine (C) at position 149 of the complementary nucleotide sequence of SEQ ID NO. 8 is substituted with thymine (T).

[0093] In the 34th aspect of the present invention, a diagnostic device for predicting the prognosis of cancer is provided, comprising: a detection unit for detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; a judgment unit for determining that the prognosis of cancer of a target individual is poor when a mutation gene is detected by the detection unit; and an output unit for outputting the judgment result of the judgment unit.

[0094] In the above embodiment, the term “diagnostic device” refers to a system, device, or apparatus, etc., comprising the detection unit, judgment unit, and output unit for predicting the prognosis of cancer. The diagnostic device is configured to perform a series of processes from the analysis of a target sample to the output of a prognosis determination result, and may include, but is not limited to, a detection unit; a judgment unit; and an output unit. Furthermore, these components may be implemented in the form of physical equipment, software modules, or a combination thereof. The cancer prognosis diagnostic device according to the present invention may include equipment that can be utilized for various clinical decision-making, such as not only being used to predict the cancer prognosis of a subject, but also determining whether to apply chemotherapy, determining whether to perform surgical treatment, predicting responsiveness to anticancer drugs, and further evaluating the prognosis after treatment.

[0095] In the above embodiment, the term “detection unit” refers to a module or means for detecting the presence of mutations on one or more genes belonging to a predefined group of genes by analyzing nucleic acids extracted from a biological sample of a target individual. The detection unit may be implemented, for example, using next-generation sequencing (NGS), polymerase chain reaction (PCR), quantitative PCR, DNA chips, or similar molecular biological techniques, but is not limited thereto. Additionally, the detection unit may include a preparation for detecting the presence of mutations on one or more genes belonging to a predefined group of genes, and the preparation for detection may include one or more selected from the group consisting of a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene, but is not limited thereto.

[0096] In the above embodiment, the term “judgment unit” refers to a module or means for performing an evaluation regarding the cancer prognosis of a target individual based on the result of the detection unit. Specifically, if a mutation is detected in the gene group, the judgment unit determines that the cancer prognosis of the target individual is poor. The judgment unit may be implemented in software, hardware, or a combination thereof, and may include a simple rule-based algorithm or a statistical or machine learning-based model.

[0097] In the above embodiment, the term “output unit” refers to a module or means that performs the function of delivering, storing, or transmitting information related to the cancer prognosis of a subject (e.g., a patient) to a user based on the analysis results derived from the judgment unit, or provides the judgment results to the user. The output means may be implemented as a display device (e.g., monitor, touchscreen, etc.), a printer, a report generation module, a computerized record linkage system via a network, etc., and, if necessary, the analysis results may be automatically transmitted to an Electronic Health Record (EHR) system or linked with a Clinical Decision Support System (CDSS) to be utilized for establishing a treatment plan. Additionally, the output unit may output the prognosis judgment results in the form of text, graphs, risk grades, etc.

[0098] In the above 34th embodiment, the 35th embodiment comprises a mutation on the gene in the detection unit in which valine (V) at position 600 in the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E) as a variant gene (p.V600E); glycine (G) at position 12 in the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid (p.G12); glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid (p.G13); glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid (p.Q61); and glutamate (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K) as a variant gene (p.E545K). A variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); a variant gene (p. C634) in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; A diagnostic device is provided that is one or more selected from the group consisting of: a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,135 (based on hg38)).

[0099] In the 36th aspect of the present invention, a method for diagnosing cancer is provided, comprising the steps of: obtaining a biological sample from a target individual; and detecting a mutation on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

[0100] In the above 36th embodiment, the 37th embodiment provides a method wherein the step of detecting the mutation is performed through a preparation comprising one or more selected from the group consisting of: a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

[0101] In the above 36th or 37th embodiment, the 38th embodiment provides a method wherein the mutation is one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, and inversion.

[0102] In any one of the above 36 to 38 embodiments, the 39th embodiment provides a method for diagnosing cancer that further comprises the step of detecting a mutation on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

[0103] In any one of the above 36th to 39th embodiments, the 40th embodiment is that the cancer is thyroid cancer, parathyroid cancer, pancreatic cancer, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, prostatic cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary gland A method is provided that is at least one selected from a group of lineages.

[0104]

[0105] The present invention relates to a combination of markers capable of predicting the prognosis of thyroid cancer with high accuracy before proceeding with surgical treatment for thyroid nodules, thereby allowing for a decision on whether to perform surgery in advance. By enabling precise prediction of the prognosis of thyroid cancer at an earlier stage, it has the effect of minimizing unnecessary surgery on thyroid nodules.

[0106] Furthermore, the effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

[0107]

[0108] FIG. 1 is a diagram showing the malignancy and mutation characteristics of indeterminate nodules (classification III and IV) according to the Korean Thyroid Imaging Reporting and Data System (KTIRADS) in accordance with an embodiment of the present invention. Here, ROM represents the risk of malignancy.

[0109]

[0110] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the invention.

[0111]

[0112] [Preparation Example] Public Database Collection

[0113] To search for genes clinically associated with thyroid cancer, various public databases were collected. By analyzing the data from the collected public databases, prognostic biomarkers for thyroid nodules were proposed, and criteria for determining importance were established to enable the detection of these biomarkers. The inventors collected and utilized various data and related literature information regarding mutations, including millions of somatic mutations detected in various tumor types, for analysis. In particular, related literature on the clinical impact of thyroid nodule-associated genes (or gene mutations) was collected to secure a database as shown in Table 1 below.

[0114] Type Collection Method DNA, SNV / INDEL, CNVMT, MSI Literature Search, Public Database RNA, Gene Fusion Literature Search, Public Database Overexpression Literature Search HKGs (Housekeeping Genes) Literature Search

[0115]

[0116] [Example 1] Data Preprocessing

[0117] Regarding the information existing in public databases, cases of germline mutations were excluded from mutation information containing information on thyroid cancer (or all solid tumors).

[0118] Item Name StandardDNA (SNV / INDEL, CNV)Tumor type compositionAt least 1 thcaSamplesAt least 1 thyroidBiomarker typeGenetic BiomarkersDiseaseThyroidCancer TypesThyroid Cancer, All Solid Tumors, THCAEvidence typePredictive (exception)Evidence directionDoes not support (exception)Variant originGermline (exception)Primary siteThyroidHistology subtype1Benign, Normal (exception)GENOMIC_MUTATION_IDCOSV (have cosmic COSV ID)Tumor originPrimary, Recurrent, NSMutation DescriptionHave Substitution infoMutation somatic statusHave somaticRNA (Gene fusion)DiseaseThyroidCancer TypeTHCATCGA Sample Id (n)At least 3 samples (no duplication)5'_GENE_NAME, 3'_GENE_NAMEUsing gene name for making fusion nameTierTier1-3Frame (exception)NA

[0119]

[0120] RNA sequencing data and in-house data were collected and utilized through public databases to confirm overexpression biomarkers and gene expression levels of HKGs.

[0121]

[0122] [Example 2] Analysis of Differentially Expressed Genes (DEGs)

[0123] To confirm biomarker overexpression in parathyroid glands and specific thyroid nodule subtypes, a patient group with overexpression (e.g., parathyroid specimens) and a control group (e.g., specimens excluding parathyroid specimens) were classified, and the presence and extent of gene RNA expression were confirmed and compared with the control group. Through the analysis of Differentially Expressed Genes (DEGs), the significance of using them as biomarkers to identify thyroid nodules was confirmed.

[0124] HKGs were identified to establish criteria for biomarker overexpression and to stabilize the quantification of gene expression values. To confirm that there was no difference in expression levels between thyroid nodules and normal tissue (control group) for the HKGs commonly presented in the two papers in Table 3 below, DEG analysis was performed individually on the two datasets, and genes with statistical significance (P < 0.05) were excluded. In addition, the coefficient of variation and standard deviation between samples within each dataset were calculated for the selected genes, and genes with a coefficient of variation of less than 30 and a standard deviation of less than 30 calculated in each dataset were selected.

[0125] Journal name CellCorrigendum to: Human housekeeping genes, revisited.Nucleic Acids ResearchHRT Atlas v1.0 database: redefining human and mouse housekeeping genes and candidate reference transcripts by mining massive RNA-seq datasets

[0126]

[0127] [Example 3] Verification of Diagnostic Criteria for Biomarker Combinations

[0128] To validate the diagnostic criteria based on the biomarker combination established in the above example, genetic testing including some biomarkers was performed. Genetic testing was conducted on thyroid nodules that underwent fine-needle aspiration biopsy, and the effect of the genetic test results on the surgical resection rate was evaluated.

[0129] Of the total 475 nodules, 278 nodules that were not genetically tested were set as a control group, and the surgical resection rates of nodules that underwent genetic testing were compared. As a result, the genetically tested group (75 / 278, 27.0%) showed a lower resection rate for all nodules than the control group (146 / 278, 52.5%). Additionally, lower surgical resection rates in the panel groups compared to the control group were also confirmed in the Bethesda III-IV groups (23.5% vs. 68.2%) and Bethesda V-VI groups (70.2% vs. 88.3%), which were classified as indeterminate nodules (see Table 4).

[0130] Groups All II III VI V VI Surgical Resection Rate Genetic Testing Group 75 / 2 78 (27.0%) 7 / 8 7 (8.0%) 23 / 10 2 (22.5%) 4 / 13 (30.8%) 4 / 9 44.4%) 36 / 48 (75.0%) Control Group 146 / 2 78 (52.5%) 15 / 88 (17.0%) 69 / 10 3 (67.0%) 6 / 7 (85.7%) 8 / 9 88.9%) 45 / 51 (88.2%) P-value <0.00 10.07 3 <0.00 10.02 20.05 20.09

[0131]

[0132] In addition, thyroid cancer-related mutations were detected in 41.7% of all specimens tested for genetic testing and in 40.0% of indeterminate nodules. Thyroid cancer-associated mutations, including V600E (Sequence No. 1), G12 (Sequence No. 2), and Q61 (Sequence No. 4), were detected (see Table 5), and it was confirmed that the surgical resection rate was higher in the groups with detected mutations, in the order of double mutation group (100%), BRAF V600E group (62.2%), RAS group (25.7%), and undetected group (18.5%) (see Table 6).

[0133]

[0134] Group Mutation Gene All(n=278)Ⅱ(n=87)Ⅲ(n=102)Ⅳ(n=13)Ⅴ(n=9)Ⅵ(n=48)noneNone*162(58.3)64(7.4)62(60.8)7(53.8)3(33.3)12(25.0)NBNRTotal32(11.5)10(11.5)18(17.6)0(0.0)0(0.0)0(0.0)EZH112(4.3)4(4.6)7(6.9)---DICER111(4.0)3(3.4)7(6 .9)---EIF1AX7(2.5)3(3.4)3(2.9)---PTEN2(0.7)-1(1.0)---RASTotal35(12.6)12(13.8)17(16.7)5(38.5)0(0.0)0(0 .0)NRAS24(8.6)10(11.5)10(9.8)3(2.3)--KRAS7(2.5)1(1.1)5(4.9)1(7.7)--HRAS4(1.4)1(1.1)2(2.0)1(7.7)--BRAF V600E BRAF V600E 45(16.2)1(1.1)4(3.9)1(7.7)6.(66.7)33(68.8)DoubleMutationTotal4(1.4)0(0.0)1(1.0)0(0.0)0(0.0)3(6.3)BRAF V600E / PIK3CA1(0.4)----1(2.1)BRAF V600E / TP531(0.4)----1(2.1)PIK3CA / TERT1(0.4)----1(2.1)EZH1 / TP531(0.4)-1(1.0)---

[0135] Molecular groupMutated genePathology of resected nodulesGroupTotalN=278ResectedN=75(27.0)GeneTotalN=278ResectedN=75(27.0)MalignantN=43(57.3)BenignN=16(21.3)UnknownN=16(21.3)No mutation162(58.3)30(18.5)None*162(58.3)30(18.5)12(8PTC, 2FTC, 2OCA)8(3FA, 1OA, 4FND)10NBNR32(12.6)4(12.5%)EZH112(4.3)1(8.3)-1(FA)-DICER111(4.0)1(9.1)-1(FA)-EIF1AX 7(2.5)2(28.6)1(FTC)1(FA)-PTEN2(0.7)----RAS35(12.6)9(25.7%)NRAS24(8.6)8(33.3%)4(2PTC, 1FTC, 1NIFTP)4(1FA, 1OA, 1FND)1KRAS7(2.5)1(25.0%)---HRAS4(1.4)1(25.0%)1(FTC)--BRAF V600E 45(16.2)28(62.2%)BRAF V600E 45(16.2)28(62.2)22(21PTC, 1DHGTC)-6 double mutation4(1.4)4(100.0)BRAF V600E / PIK3CA1(0.4)1(100.0)1(PTC)--BRAF V600E / TP531(0.4)1(100.0)1(PTC)-PIK3CA / TERT1(0.4)1(100.0)1(PTC)--EZH1 / TP531(0.4)1(100.0)-1(OA)

[0136]

[0137] Additionally, the risk of malignancy was evaluated based on the results of gene panel testing and ultrasound characteristics, and it was confirmed that in all K-TIRADS categories, the risk of malignancy was higher in undetermined nodules with mutations than in undetermined nodules without mutations (see Fig. 1).

[0138] Referring to Figure 1, the risk of malignancy (ROM) of indeterminate nodules without mutations was 11.1%, 2.8%, and 0.0% in K-TIRADS 3, 4, and 5, respectively, while nodules with mutations showed higher risk of malignancy at 28.6%, 6.7%, and 22.2%.

[0139] Similarly, by confirming that the risk of malignancy in unidentified nodule specimens with detected mutations among specimens that underwent gene panel testing and surgical resection is higher than that of specimens without detection, it can be confirmed that the detection of biomarkers through gene panel testing is effective in reducing unnecessary surgeries.

[0140] Foregoing, specific parts of the present invention have been described in detail. It is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

[0141]

[0142] Sequence No. 1: BRAF partial sequence

[0143] MAALSGGGGGGGAEPGQALFNGDMEPEAGAGAAGAAASSAADPAIPEEVWNIKQMIKLTQEH

[0144] IEALLDKFGGEHNPPSIYLEAYEEYTSKLDALQQREQQLLESLGNGTDFSVSSSASMDTV

[0145] TSSSSSSLSVLPSSLSVFQNPTDVARSNPKSPQKPIVRVFLPNKQRTVVPARCGVTVRDS

[0146] LKKALMRGLIPECCAVYRIQDGEKKPIGWDTDISWLTGEELHVEVLENVPLTTHNFVRK

[0147] TFFTLAFCDFCRKLLFQGFRCQTCGYKFHQRCSTEVPLMCVNYDQLDLLFVSKFFEHHPI

[0148] PQEEASLAETALTSGSSPSAPASDSIGPQILTSPSPSKSIPIPQPFRPADEDHRNQFGQR

[0149] DRSSSAPNVHINTIEPVNIDDLIRDQGFRGDGGSTTGLSATPPASLPGSLTNVKALQKSP

[0150] GPQRERKSSSSSEDRNRMKTLGRRDSSDDWEIPDGQITVGQRIGSGSFGTVYKGKWHGDV

[0151] AVKMLNVTAPTPQQLQAFKNEVGVLRKTRHVNILLFMGYSTKPQLAIVTQWCEGSSLYHH

[0152] LHIIETKFEMIKLIDIARQTAQGMDYLHAKSIIHRDLKSNNIFLHEDLTVKIGDFGLATV

[0153] KSRWSGSHQFEQLSGSILWMAPEVIRMQDKNPYSFQSDVYAFGIVLYELMTGQLPYSNIN

[0154] NRDQIIFMVGRGYLSPDLSKVRSNCPKAMKRLMAECLKKKRDERPLFPQILASIELLARS

[0155] LPKIHRSASEPSLNRAGFQTEDFSLYACASPKTPIQAGGYGAFPVH

[0156] 서열번호 2 : KRAS partial sequence

[0157] MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAG

[0158] QEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKDSEDVPMVLVGNKCDL

[0159] PSRTVDTKQAQDLARSYGIPFIETSAKTRQRVEDAFYTLVREIRQYRLKKISKEEKTPGC

[0160] VKIKKCIIM

[0161] 서열번호 3 : HRAS partial sequence

[0162] MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAG

[0163] QEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKDSDDVPMVLVGNKCDL

[0164] AARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIRQHKLRKLNPPDESGPG

[0165] CMSCKCVLS

[0166] 서열번호 4 : NRAS partial sequence

[0167] MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAG

[0168] QEEYSAMRDQYMRTGEGFLCVFAINNSKSFADINLYREQIKRVKDSDDVPMVLVGNKCDL

[0169] PTRTVDTKQAHELAKSYGIPFIETSAKTRQGVEDAFYTLVREIRQYRMKKLNSSDDGTQG

[0170] CMGLPCVVM

[0171] 서열번호 5 : PIK3CA partial sequence

[0172] MPPRPSSGELWGIHLMPPRILVECLLPNGMIVTLECLREATLITIKHELFKEARKYPLHQ

[0173] LLQDESSYIFVSVTQEAEREEFFDETRRLCDLRLFQPFLKVIEPVGNREEKILNREIGFA

[0174] IGMPVCEFDMVKDPEVQDFRRNILNVCKEAVDLRDLNSPHSRAMYVYPPNVESSPELPKH

[0175] IYNKLDKGQIIVVIWVIVSPNNDKQKYTLKINHDCVPEQVIAEAIRKKTRSMLLSSEQLK

[0176] LCVLEYQGKYILKVCGCDEYFLEKYPLSQYKYIRSCIMLGRMPNLMLMAKESLYSQLPMD

[0177] CFTMPSYSRRISTATPYMNGETSTKSLWVINSALRIKILCATYVNVNIRDIDKIYVRTGI

[0178] YHGGEPLCDNVNTQRVPCSNPRWNEWLNYDIYIPDLPRAARLCLSICSVKGRKGAKEEHC

[0179] PLAWGNINLFDYTDTLVSGKMALNLWPVPHGLEDLLNPIGVTGSNPNKETPCLELEFDWF

[0180] SSVVKFPDMSVIEEHANWSVSREAGFSYSHAGLSNRLARDNELRENDKEQLKAISTRDPL

[0181] SEITEQEKDFLWSHRHYCVTIPEILPKLLLSVKWNSRDEVAQMYCLVKDWPPIKPEQAME

[0182] LLDCNYPDPMVRGFAVRCLEKYLTDDKLSQYLIQLVQVLKYEQYLDNLLVRFLLKKALTN

[0183] QRIGHFFFWHLKSEMHNKTVSQRFGLLLESYCRACGMYLKHLNRQVEAMEKLINLTDILK

[0184] QEKKDETQKVQMKFLVEQMRRPDFMDLQGFLSPLNPAHQLGNRLRLEECRIMSSAKRPLW

[0185] LNWENPDIMSELLFQNNEIIFKNGDDLRQDMLTLQIIRIMENIWQNQGLDLRMLPYGCLS

[0186] IGDCVGLIEVVRNSHTIMQIQCCGGLKGALQFNSHTLHQWLKDKNKGEIYDAAIDLFTRS

[0187] CAGYCVATFILGIGDRHNSNIVMKDDGQLFHIDFGHFLDHKKKKFGYKRERVPFVLTQDF

[0188] LIVISKGAQECTKTREFERFQEMCYKAYLAIRQHANLFINLFSMMLGSGMPELQSFDDIA

[0189] YIRKTLALDKTEQEALEYFMKQMNDAHHGGWTTKMDWIFHTIKQHALN

[0190] sökðiboldeh 6 : RET partial sequence

[0191] MAKATSGAAGLRLLLLLLLPLLGKVALGLYFSRDAYWEKLYVDQAAGTPLLYVHALRDAP

[0192] EEVPSFRLGQHLYGTYRTRLHENNWICIQEDTGLLYLNRSLDHSSWEKLSVRNRGFPLLT

[0193] VYLKVFLSPTSLREGECQWPGCARVYFSFFNTSFPACSSLKPRELCFPETRPSFRIRENR

[0194] PPGTHFQFRLLPVQFLCPNISVAYRLLEGEGLPFRCAPDSLEVSTRWALDREQREKYELV

[0195] AVCTVHAGAREEVVMVPPFVTVYEDDDSAPTFPAGVDTASAVVEFKRKEDTVVATLRVFD

[0196] ADVVPASGELVRRYTSTLLPGDTWAQQTFRVEHWPNETSVQANGSFVRATVHDYRLVLNR

[0197] NLSISENRTMQLAVLVNDSDFQGPAGVLLLHFNVSVLPVSLHLPSTYSLSVSRRARRFA

[0198] QIGKVCVENCQAFSGINVQYKLHSSGANCSTLGVVTSAEDTSGILFVNDTKALRRPKCAE

[0199] LHYMVVADTQQTSRQAQAQLLVTVEGSYVAEEAGCPLSCAVSKRRLECEECGGLGSPTGR

[0200] CEWRQDGDGKGITRNFSTCSPSTKTCPDGHCDVVETQDINICPQDCLRGSIVGGHEPGEPR

[0201] GIKAGYGTCNCFPEEEKCFCEPEDIQDPLCDELCRTVIAAVLFSFIVSVLLSAFCIHCY

[0202] HKFAHKPPISSAEMTFRRPAQAFPVSYSSSGARRPSLDSMENQVSVDAFKILEDPKWEFP

[0203] RKNLVLGKTLGEGEFGKVVKATAFHLKGRAGYTTVAVKMLKENASPSELRDLLSEFNVLK

[0204] QVNHPHVIKLYGACSQDGPLLLIVEYAKYGSLRGFLRESRKVGPGYLGSGGSRNSSSLDH

[0205] PDERALTMGDLISFAWQISQGMQYLAEMKLVHRDLAARNILVAEGRKMKISDFGLSRDVY

[0206] EEDSYVKRSQGRIPVKWMAIESLFDHIYTTQSDVWSFGVLLWEIVTLGGNPYPGIPPERL

[0207] FNLLKTGHRMERPDNCSEEMYRLMLQCWKQEPDKRPVFADISKDLEKMMVKRRDYLDLAA

[0208] STPSDSLIYDDGLSEEETPLVDCNNAPLPRALPSTWIENKLYGMSDPNWPGESPVPLTRA

[0209] DGTNTGFPRYPNDSVYANWMLSPSAAKLMDTFDS

[0210] 서열번호 7 : TP53 partial sequence

[0211] MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGP

[0212] DEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAK

[0213] SVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHE

[0214] RCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNS

[0215] SCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELP

[0216] PGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPG

[0217] GSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD

[0218] 서열번호 8 : TERT partial sequence, chr5:1294987-1295135

[0219] CATCGCGGGGGTGGCCGGGGCCAGGGCTTCCCACGTGCGCAGCAGGACGC

[0220] AGCGCTGCCTGAAACTCGCGCCGCGAGGAGAGGGCGGGGCCGCGGAAAGG

[0221] AAGGGGAGGGGCTGGGAGGGCCCGGAGGGGGCTGGGCCGGGGACCCGGG

Claims

1. A composition for predicting the prognosis of cancer, comprising a preparation for detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

2. In Paragraph 1, The above-mentioned preparation is a composition comprising one or more selected from the group consisting of: a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

3. In Paragraph 1, A composition wherein the above mutation is one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, and inversion.

4. In Paragraph 1, The above BRAF mutation is, A composition characterized by being a variant gene (p.V600E) in which valine (V) at position 600 of the amino acid sequence of SEQ ID NO. 1 is substituted with glutamic acid (E).

5. In Paragraph 1, The above KRAS mutation is, A composition characterized by being a variant gene (p.G12) in which the glycine (G) at position 12 of the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid.

6. In Paragraph 1, The above HRAS mutation is, A composition characterized by being a variant gene (p.G13) in which the glycine (G) at position 13 of the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid.

7. In Paragraph 1, The above NRAS mutation is, A composition characterized by being a variant gene (p.Q61) in which glutamine (Q) at position 61 of the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid.

8. In Paragraph 1, The above PIK3CA mutation is, A variant gene (p.E545K) in which glutamic acid (E) at position 545 of the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K); and A composition characterized by being one or more selected from the group consisting of a variant gene (p. H1047R) in which histidine (H) at position 1047 in the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R).

9. In Paragraph 1, The above RET mutation is, A variant gene (p. C634) in which the cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; and A composition characterized by being one or more selected from the group consisting of a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T).

10. In Paragraph 1, The above TP53 mutation is, A variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); and A composition characterized by being one or more selected from the group consisting of: a variant gene (p. R273) in which the arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid.

11. In Paragraph 1, The above TERT mutation is, A promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and A composition characterized by being one or more selected from the group consisting of a promoter variant gene (chr5: 1,295,135 (based on hg38)) in which cytosine (C), the complementary base sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T).

12. In Paragraph 1, The above composition further comprises a preparation for detecting mutations on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

13. In Paragraph 12, The above RET mutation is, A composition in which the RET gene appears in a fused form with CCDC6.

14. In Paragraph 12, The above BRAF mutation is, A composition in which the BRAF gene appears in a fused form with AKAP9.

15. In Paragraph 12, The above NTRK1 mutation is, A composition in which the NTRK1 gene appears in a fused form with TPM3.

16. In Paragraph 1, A composition comprising at least one selected from the group consisting of the above cancers: thyroid cancer, parathyroid cancer, pancreatic cancer, stomach cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, pro-anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma.

17. A kit for predicting the prognosis of cancer comprising a composition of any one of claims 1 to 16.

18. A method for providing information regarding the prognosis prediction of cancer, comprising the step of detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT in a biological sample isolated from a target individual.

19. In Paragraph 18, The above method further comprises the step of detecting a mutation on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

20. In Paragraph 18, The above mutant gene is a variant gene (p.V600E) in which valine (V) at position 600 in the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E); a variant gene (p.G12) in which glycine (G) at position 12 in the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid; a variant gene (p.G13) in which glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid; a variant gene (p.Q61) in which glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid; a variant gene (p.E545K) in which glutamate (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K); a variant gene (p.H1047R) in which histidine (H) at position 1047 in the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); A variant gene in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid (p. C634); a variant gene in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T) (p. M918T); a variant gene in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H) (p. R175H); a variant gene in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid (p. R273); a promoter variant gene in which cytosine (C), the complementary base sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and if one or more are selected from the group consisting of a promoter variant gene (chr5: 1,295,135 (based on hg38)) in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T), A method characterized by predicting that the above-mentioned entity will have a poor prognosis for cancer.

21. In Paragraph 19, The above-mentioned mutant gene, If one or more are selected from the group consisting of: a fusion gene in which the RET gene is fused with the CCDC6 gene; a fusion gene in which the BRAF gene is fused with the AKAP9 gene; and a fusion gene in which the NTRK1 gene is fused with the TPM3 gene, A method characterized by predicting that the above-mentioned entity will have a poor prognosis for cancer.

22. In Paragraph 18, The above cancer is at least one selected from the group consisting of thyroid cancer, parathyroid cancer, pancreatic cancer, stomach cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, pro-anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma, a method.

23. Use of an agent for cancer prognosis prediction to detect mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

24. In Paragraph 23, The above preparation is for use in predicting cancer prognosis, comprising one or more selected from the group consisting of: a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

25. In Paragraph 23, The above mutation is one or more mutations selected from a group consisting of single nucleotide variation (SNV), frame shift mutation, insertion / deletion mutation (Indel), copy number variation (CNV), deletion, and inversion, for use in predicting cancer prognosis.

26. In Paragraph 23, The above BRAF mutation is a variant gene (p.V600E) in which valine (V) at position 600 of the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E), and is used for cancer prognosis prediction.

27. In Paragraph 23, The above KRAS mutation is a variant gene (p.G12) in which glycine (G) at position 12 of the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid, and is used for cancer prognosis prediction.

28. In Paragraph 23, The above HRAS mutation is a variant gene (p.G13) in which glycine (G) at position 13 of the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid, and is used for cancer prognosis prediction.

29. In Paragraph 23, The above NRAS mutation is a variant gene (p.Q61) in which glutamine (Q) at position 61 of the amino acid sequence of SEQ No. 4 is substituted with another amino acid, used for cancer prognosis prediction.

30. In Paragraph 23, The above PIK3CA mutation is a variant gene (p.E545K) in which glutamic acid (E) at position 545 of the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K); and One or more selected from the group consisting of a variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); for use in predicting cancer prognosis.

31. In Paragraph 23, The above RET mutation is a variant gene (p. C634) in which the cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; and One or more selected from the group consisting of a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T). Uses for predicting cancer prognosis.

32. In Paragraph 23, The above TP53 mutation is a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); and One or more selected from the group consisting of a variant gene (p. R273) in which the arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; and used for predicting cancer prognosis.

33. In Paragraph 23, The above TERT mutation is a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and One or more selected from the group consisting of a promoter variant gene (chr5: 1,295,135 (based on hg38)) in which cytosine (C), the complementary nucleotide sequence at position 149 of sequence number 8, is substituted with thymine (T). Uses for predicting cancer prognosis.

34. As a diagnostic device for predicting the prognosis of cancer, A detection unit for detecting mutations on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; A judgment unit that determines that the prognosis of cancer in the target individual will be poor when a mutant gene is detected by the above detection unit; and A diagnostic device comprising: an output unit that outputs the judgment result of the above-mentioned judgment unit.

35. In Paragraph 34, The mutations on the genes in the above detection unit are a variant gene (p.V600E) in which valine (V) at position 600 in the amino acid sequence of SEQ ID NO. 1 is substituted with glutamate (E); a variant gene (p.G12) in which glycine (G) at position 12 in the amino acid sequence of SEQ ID NO. 2 is substituted with another amino acid; a variant gene (p.G13) in which glycine (G) at position 13 in the amino acid sequence of SEQ ID NO. 3 is substituted with another amino acid; a variant gene (p.Q61) in which glutamine (Q) at position 61 in the amino acid sequence of SEQ ID NO. 4 is substituted with another amino acid; and a variant gene (p.E545K) in which glutamate (E) at position 545 in the amino acid sequence of SEQ ID NO. 5 is substituted with lysine (K). A variant gene (p. H1047R) in which histidine (H) at position 1047 of the amino acid sequence of SEQ ID NO. 5 is substituted with arginine (R); a variant gene (p. C634) in which cysteine ​​(C) at position 634 of the amino acid sequence of SEQ ID NO. 6 is substituted with another amino acid; a variant gene (p. M918T) in which methionine (M) at position 918 of the amino acid sequence of SEQ ID NO. 6 is substituted with tyrosine (T); a variant gene (p. R175H) in which arginine (R) at position 175 of the amino acid sequence of SEQ ID NO. 7 is substituted with histidine (H); a variant gene (p. R273) in which arginine (R) at position 273 of the amino acid sequence of SEQ ID NO. 7 is substituted with another amino acid; A diagnostic device comprising one or more selected from the group consisting of: a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 127 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,113 (based on hg38)); and a promoter variant gene in which cytosine (C), the complementary nucleotide sequence at position 149 of SEQ ID NO. 8, is substituted with thymine (T) (chr5: 1,295,135 (based on hg38)).

36. A step of obtaining a biological sample from a target individual; and A method for diagnosing cancer comprising the step of detecting a mutation on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT.

37. In Paragraph 36, A method wherein the step of detecting the mutation is performed using a preparation comprising one or more selected from the group consisting of: a primer or probe that specifically binds to a mutation site on one or more genes selected from the group consisting of BRAF, KRAS, HRAS, NRAS, PIK3CA, RET, TP53, and TERT; and an antibody that specifically binds to a protein encoded by said gene.

38. In Paragraph 36, A method wherein the mutation is one or more mutations selected from the group consisting of single nucleotide variation (SNV), frame shift variation, insertion / deletion variation (Indel), copy number variation (CNV), deletion, and inversion.

39. In Paragraph 36, The method for diagnosing the above cancer further comprises the step of detecting a mutation on one or more genes selected from the group consisting of RET, BRAF, and NTRK1.

40. In Paragraph 36, The above cancer is at least one selected from the group consisting of thyroid cancer, parathyroid cancer, pancreatic cancer, stomach cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, pro-anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma.

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