Methods for sensitizing to chemotherapy drugs, their active ingredient composition and use

Abstract

Procedures for sensitization to chemotherapy drugs, including: Transfecting an antisense oligonucleotide into a cancer cell to form a transfected cancer cell to block the cancer cell's miR-1976 during future treatment; Delivering a treatment with a cancer drug to the transfected cancer cell whose miR-1976 is blocked by the antisense oligonucleotide; and Offering an increased degree of sensitivity to the cancer drug through the transfected cancer cell during treatment with a cancer drug, as the miR-1976 of the cancer cell is blocked.

Description

BACKGROUND OF THE INVENTION 1. Field of the invention

[0001] The present invention relates to a method for sensitizing chemotherapy drugs, an active ingredient composition thereof (or a test reagent), and a method of use. In particular, the present invention relates to increasing the sensitivity of a method for sensitizing chemotherapy drugs, an active ingredient composition thereof (or a test reagent), and a method of use. More specifically, the present invention relates to the method for sensitizing chemotherapy drugs, the active ingredient composition thereof (or the test reagent), and the method of use, which utilizes the inhibition of miR-1976 nucleic acids to sensitize chemotherapy drugs for cancers. 2. Description of the state of the art

[0002] Cancers in humans generally include brain cancer, nasopharyngeal cancer (NPC), oral cancer, laryngeal cancer, esophageal cancer, stomach cancer, liver cancer, colon cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, cervical cancer, germ cell cancer, skin cancer, osteosarcoma, blood cancers (including lymphoma, leukemia, and multiple myeloma), and so on. Pancreatic cancer, for example, is one of the most common cancers worldwide. A 2017 announcement by the Taiwanese Ministry of Health and Welfare shows that malignant tumors have been among the top ten causes of death in recent years, with pancreatic cancer ranking sixth among them. Pancreatic cancer is most common in industrialized countries, yet the prognosis for pancreatic cancer is usually quite poor, with 1-year and 5-year survival rates of only 25% and 5%, respectively.In some cases detected early, the 5-year survival rate can rise to 20%, but survival cannot be efficiently controlled.

[0003] US patent no. 8,969,315, entitled “ENHANCEMENT OF PLACENTAL STEM CELL POTENCY USING MODULATORY RNA MOLECULES”, only discloses miR-1976 (i.e., microRNA-1976) and apparently does not teach any inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0004] Another US patent application, publication number 2014 / 0080894 and titled “ENHANCED BIODISTRIBUTION OF OLGOMERS”, only discloses miR-1976 (i.e., microRNA-1976) and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0005] Another US patent application, publication number 2018 / 0237772 and titled “HYBRID tRNA / pre-miRNA MOLECULES AND METHODS OF USE”, only discloses miR-1976 (i.e., microRNA-1976) and apparently does not teach any inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0006] Another US patent, No. 8,021,840, entitled “DIAGNOSTIC MARKER FOR INTERFERON RESPONSIVENESS IN MULTIPLE SCLEROSIS”, only discloses the protein gene XAF1 and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0007] Another US patent application with publication number 2007 / 0218493 and the title “DIAGNOSTIC MARKER FOR INTERFERON RESPONSIVENESS IN MULTIPLE SCLEROSIS” only discloses the protein gene XAF1 and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0008] Another US patent application with publication number 2009 / 0215895 and the title “THERAPEUTIC AND CARRIER MOLECULES” only discloses the protein gene XAF1 and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0009] Another US patent application with publication number 2017 / 0003277 and the title “BIOLOGICAL CHARATERIZATION OF A GLATIRAMER ACETATE RELATED DRUG PRODUCT USING MAMMALIAN AND HUMAN CELLS” only discloses the protein gene XAF1 and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0010] Another US patent application with publication number 2018 / 0369303 and the title “ONCOLYTIC RHABDOVIRUS” only discloses the protein gene XAF1 and apparently does not teach an inhibition technique for miR-1976 nucleic acids and possibilities of associated apoptosis of cancer cells.

[0011] The journal “Chinese Journal of Cancer” with LIU et al. “Chin J Cancer (2015) 34:44” reveals under the title “miR-506: a regulator of chemo-sensitivity through suppression of the RAD51-homologous recombination axis” the increased chemosensitivity to cisplatin and olaparib by miR-506 via RA-D52 suppression in ovarian carcinoma models.

[0012] The journal “MOLECULAR MEDICINE REPORTS” with XIONG et al. “MOLECULAR MEDICINE REPORTS 12: 5019-5025, 2015” reveals under the title “MicroRNA-197 reverses the drug resistance of fluorouracil-induced SGC7901 cells by targeting mitogen-activated protein kinase 1” that in DGC7901 / 5-FU gastric cancer cells resistant to fluorouracil, overexpression of miR-197 restores sensitivity to fluorouracil.

[0013] However, there is a need to provide an inhibition technique for miR-1976 nucleic acids and possibilities for associated apoptosis of cancer cells. The patents and patent application publications mentioned above are incorporated herein by reference for purposes including, but not limited to, providing background information on the present invention and illustrating the prior art. SUMMARY OF THE INVENTION

[0014] The main objective of this invention is to provide a method for sensitizing cells to chemotherapy drugs, an active ingredient composition thereof (or a test reagent), and a method of use. A cancer cell is transfected (or treated) to form a cancer cell transfected with an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) to block the cancer cell's miR-1976 during future treatment. The transfected cancer cell is treated with a cancer drug (or an anticancer drug combination) such that the antisense oligonucleotide blocks the miR-1976 of the transfected cancer cell. Upon treatment with a cancer drug, the transfected cancer cell exhibits an increased degree of sensitivity to the cancer drug because its miR-1976 is blocked.The method for sensitizing to chemotherapy drugs, the active ingredient composition (or the test reagent) and the use of the present invention are advantageously successful in providing an increased degree of sensitivity to various cancer drugs.

[0015] The method for sensitizing to chemotherapy drugs according to one aspect of the present invention includes: Transfecting an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) into a cancer cell to form a transfected cancer cell to block the cancer cell's miR-1976 in future treatment; Delivering treatment with a cancer drug (or an anticancer drug combination) to the transfected cancer cell whose miR-1976 is blocked by the antisense oligonucleotide; and Offering an increased degree of sensitivity to the cancer drug through the transfected cancer cell during treatment with a cancer drug, as the miR-1976 of the cancer cell is blocked. The method for sensitizing to chemotherapy drugs according to one aspect of the present invention further includes: Searching for at least one molecular target in the cancer cell; Screening at least one microRNA-binding target within a predetermined cell; Providing a target-binding RNA fragment to be transcribed or extracted into the microRNA-binding target; Providing a sequential analysis for the target-binding RNA fragment and determining a target gene and its corresponding position to obtain a variety of miR-1976-binding targets; and Searching for an XAF1 gene with the multitude of miR-1976 binding targets and providing an antisense oligonucleotide transfection procedure (e.g., siRNA-1976 or antagomir-1976 transfection procedure) for the cancer cell to block the cancer cell's miR-1976.

[0016] In a separate aspect of the present invention, the at least one microRNA-binding target is screened by cloning specific genes.

[0017] In a further separate aspect of the present invention, the target gene and its corresponding position are obtained by blast analysis.

[0018] In yet another separate aspect of the present invention, an expression of miR-1976 is obtained by microarray analysis or qPCR analysis.

[0019] The active ingredient composition for sensitization to chemotherapy drugs according to one aspect of the present invention includes: an antisense oligonucleotide provided to treat a cancer cell with an antisense oligonucleotide transfection procedure (e.g., siRNA-1976 or antagomir-1976 transfection procedure) to create a transfected cancer cell to block the cancer cell's miR-1976 during future treatment; and a cancer drug (or an anticancer drug combination) that is delivered to the transfected cancer cell, administered as a combination to increase sensitivity; wherein the cancer drug is provided to treat the transfected cancer cell whose miR-1976 is blocked, the miR-1976-blocked cancer cell offering an increased degree of sensitivity to the cancer drug.

[0020] In a separate aspect of the present invention, the combination for increasing sensitivity causes a reduction of miR-1976 in the extracellular vesicle.

[0021] In a further separate aspect of the present invention, the combination for increasing sensitivity causes an increase in cancer cell death or a reduction in cancer cell growth.

[0022] In a further separate aspect of the present invention, the combination for increasing sensitivity causes an increase in a cancer cell apoptosis marker.

[0023] In yet another separate aspect of the present invention, the cancer cell apoptosis marker is selected from cleaved PARP.

[0024] In yet another separate aspect of the present invention, the cancer cell apoptosis marker corresponds to an XAF1 gene of the cancer cell.

[0025] The method for sensitizing to chemotherapy drugs according to one aspect of the present invention includes: Transfecting an antisense oligonucleotide (e.g., siRNA-1976 or antagomir-1976) into a pancreatic cancer cell to form a transfected pancreatic cancer cell to block the pancreatic cancer cell's miR-1976 in future treatment;

[0026] Delivering treatment with a pancreatic cancer drug (or an anticancer drug combination) to the transfected pancreatic cancer cell whose miR-1976 is blocked by the antisense oligonucleotide; and

[0027] Offering an increased degree of sensitivity to the pancreatic cancer drug by the transfected pancreatic cancer cell during treatment with a pancreatic cancer drug, as the miR-1976 of the pancreatic cancer cell is blocked.

[0028] The method for sensitizing to chemotherapy drugs according to one aspect of the present invention further includes: Search for at least one molecular target in the pancreatic cancer cell; Screening for at least one microRNA-binding target within a predetermined pancreatic cell; Providing a target-binding RNA fragment to be transcribed or extracted into the microRNA-binding target; Providing a sequential analysis for the target-binding RNA fragment and determining a target gene and its corresponding position to obtain a variety of miR-1976-binding targets; and

[0029] Searching for an XAF1 gene with a multitude of miR-1976 binding targets and providing an antisense oligonucleotide transfection procedure (e.g., siRNA-1976 or antagomir-1976 transfection procedure) for the pancreatic cancer cell to block the pancreatic cancer cell's miR-1976.

[0030] In a separate aspect of the present invention, the at least one microRNA-binding target is screened by cloning specific genes.

[0031] In a further separate aspect of the present invention, the target gene and its corresponding position are obtained by blast analysis.

[0032] In yet another separate aspect of the present invention, expression of miR-1976 is obtained by microarray analysis, qPCR analysis or RNA sequence analysis.

[0033] The active ingredient composition for sensitization to chemotherapy drugs according to one aspect of the present invention includes: an antisense oligonucleotide provided to treat a pancreatic cancer cell with an antisense oligonucleotide transfection procedure (e.g., siRNA-1976 or antagomir-1976 transfection procedure) to form a transfected pancreatic cancer cell to block the miR-1976 of the pancreatic cancer cell in a future treatment; and a pancreatic cancer drug (or an anticancer drug combination) that is provided to the transfected pancreatic cancer cell, which are administered as a combination to increase sensitivity; wherein the pancreatic cancer drug is provided to treat the transfected pancreatic cancer cell whose miR-1976 is blocked, the miR-1976-blocked pancreatic cancer cell offering an increased degree of sensitivity to the pancreatic cancer drug.

[0034] In a separate aspect of the present invention, the combination for increasing sensitivity causes a reduction of miR-1976 in the extracellular vesicle.

[0035] In a further separate aspect of the present invention, the combination for increasing sensitivity causes an increase in cancer cell death or a reduction in cancer cell growth.

[0036] In a further separate aspect of the present invention, the combination for increasing sensitivity causes an increase in a pancreatic cancer cell apoptosis marker.

[0037] In yet another separate aspect of the present invention, the pancreatic cancer cell apoptosis marker is selected from cleaved PARP.

[0038] In yet another separate aspect of the present invention, the pancreatic cancer cell apoptosis marker corresponds to an XAF1 gene of the cancer cell.

[0039] A further scope of applicability of the present invention will become apparent from the detailed description given below. It should be understood, however, that the detailed description and specific examples, even if they represent preferred embodiments of the invention, are given for illustrative purposes only, since various aspects will become obvious to a person skilled in the art from this detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The present invention will be better understood with reference to the detailed description given below and the accompanying drawings, which are provided for illustrative purposes only and therefore do not limit the present invention, and of which: Fig. 1 is a flowchart of a method for sensitization to chemotherapy drugs according to a preferred embodiment of the present invention. Fig. 1A and Fig. 1B Diagrams of microRNA expression released in the extracellular vesicle and intracellularly, obtained by microarray analysis and qPCR analysis, which were applied in the sensitization process for chemotherapy drugs according to the preferred embodiment of the present invention. Fig. 2A and Fig. 2B is a diagram of the miR-1976 binding target obtained by cloning specific genes and a diagram of potential target genes (related to cancer cell apoptosis) compared with bioinformatics (XAF1) applied in the method for sensitizing to chemotherapy drugs according to the preferred embodiment of the present invention. Fig. 3A to 3C are diagrams of miR-1976 expression obtained by qPCR analysis and Western blot analysis, which were used in the sensitization process for chemotherapy drugs according to the preferred embodiment of the present invention. Fig. 4A and Fig. 4B Diagrams of cell apoptosis of BxPC-3 and AsPC-1 pancreatic cancer cells with AntagomiR-1976 transfection are obtained by Western blot analysis, which was applied in the method for sensitization to chemotherapy drugs according to the preferred embodiment of the present invention. Fig. 5A to 5D diagrams of the cancer cell viability of MIA Capa-2, BxPC-3 and AsPC-1 pancreatic cancer cells with siRNA-1976 transfection, which were used in the method for sensitization to chemotherapy drugs according to the preferred embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0041] It is pointed out that a method for sensitizing to chemotherapy drugs, an active ingredient composition thereof (or a test reagent) and a method of use according to the preferred embodiment of the present invention can be applied to various types of cancer, including brain cancer, nasopharyngeal cancer (NPC), oral cancer, laryngeal cancer, esophageal cancer, stomach cancer, liver cancer, colon cancer, pancreatic cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, cervical cancer, germ cell cancer, skin cancer, osteosarcoma cancer, blood cancer (including lymphoma cancer, leukemia and multiple myeloma), which do not limit the present invention.

[0042] A method for sensitizing chemotherapy drugs, an active ingredient composition thereof (or a test reagent) and a method of use according to a further preferred embodiment of the present invention provide, for example, a sensitizing agent for chemotherapy drugs to increase a degree of sensitivity to cancer drugs which can be used in chemotherapy treatment at a lower dose prescription, with an increase in the therapeutic effect.

[0043] The method for sensitizing chemotherapy drugs, the active ingredient composition thereof (or the test reagent), and the method of use according to the preferred embodiment of the present invention are provided, for example, to block cell physiological effects of miR-1976-, to achieve a sensitization effect of cancer cells to chemotherapy drugs, and to inhibit the growth of cancer cells. In addition, the microRNA is used to screen molecular targets of cells.

[0044] The method for sensitizing chemotherapy drugs, the active ingredient composition thereof (or the test reagent), and the method of use according to the preferred embodiment of the present invention are provided, for example, by a technology for cloning specific genes. In addition, the method for sensitizing chemotherapy drugs, the active ingredient composition thereof (or the test reagent), and the method of use according to the preferred embodiment of the present invention are provided to form a test reagent for testing the amount of miR-1976 in blood samples after injection of a cancer drug.

[0045] Fig. Figure 1 shows a flowchart of a method for sensitization to chemotherapy drugs according to a preferred embodiment of the present invention. Referring now to Fig. 1 includes, for example, the method for sensitization to chemotherapy drugs according to the preferred embodiment of the present invention: Transfecting an antisense oligonucleotide (e.g., siRNA-1976 or anthagomir-1976) into a cancer cell 1 (e.g., pancreatic cancer cell or other cancer cell) using an antisense oligonucleotide transfection procedure 2 (e.g., siRNA-1976 or anthagomir-1976 transfection procedure) to form a transfected cancer cell 10 to block the miR-1976 of cancer cell 1 in a future treatment; Delivering a treatment with a cancer drug (or an anticancer drug combination) 3 to the transfected cancer cell 10 whose miR-1976 is blocked by the antisense oligonucleotide; and Offering an increased degree of sensitivity to the cancer drug 3 by the transfected cancer cell 10 as miR-1976-blocked cancer cell 11 during treatment with a cancer drug, since the miR-1976 of the cancer cell 1 is blocked.

[0046] Still referring to Fig. 1 further includes the method for sensitization to chemotherapy drugs according to one aspect of the present invention: Search for at least one molecular target in the cancer cell 1; Screening at least one microRNA-binding target within a predetermined cell; Providing a target-binding RNA fragment to be transcribed or extracted into the microRNA-binding target; Providing a sequential analysis for the target-binding RNA fragment and determining a target gene and its corresponding position to obtain a variety of miR-1976-binding targets; and Searching for an XAF1 gene with the multitude of miR-1976 binding targets and providing the antisense oligonucleotide transfection procedure 2 (e.g. siRNA-1976 or antagomir-1976 transfection procedure) to cancer cell 1 to block the miR-1976 of cancer cell 1.

[0047] With continuous reference to Fig. 1 includes the antisense oligonucleotide transfection procedure 2 in a preferred embodiment, for example, an siRNA-1976 transfection procedure of pancreatic cancer (or other cancers) or an antagomir-1976 transfection procedure of pancreatic cancer (or other cancers).

[0048] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a preferred embodiment is selected, for example, from various antimetabolites, including methotrexate, pemetrexed, cytarabine, 5-fluorouracil, capecitabine, gemcitabine, 6-mercaptopurine, azathioprine, fludarabine, cladribine, hydroxyurea or the like.

[0049] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various alkylating agents, including cyclophosphamide, ifosfamide, chlorambucil, melphalan, temozolomide, carmustine, lomustine, streptozocin, busulfan, procarbazine, cisplatin, carboplatin, oxaliplatin or the like.

[0050] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various topoisomerase inhibitors, including irinotecan, topotecan, etoposide, doxorubicin or the like.

[0051] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various mitotic inhibitors, including vincristine, vinblastine, vinorelbine, docetaxel, paclitaxel, eribulin, ixabepilone, epothilone or the like.

[0052] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various antitumor antibiotics, including bleomycin, actinomycin D, doxorubicin, daunorubicin, idarubicin, mitomycin or the like.

[0053] With continuous reference to Fig. 1 the cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various protein kinase inhibitors, including imatinib, dasatinib, nilotinib, erlotinib, gefitinib, afatinib, osimertinib, alectinib, crizotinib, dabrafenib, encorafenib, vemurafenib, trametinib, ibrutinib, ruxolitinib, palbociclib or the like.

[0054] With continuous reference to Fig. 1 The cancer drug (or anticancer drug combination) 3 in a further preferred embodiment is selected, for example, from various monoclonal antibody drugs against tumors or tumor receptors, or can be selected from L-asparaginase, bortezomib, carfilzomib, ixazomib, olaparib or the like.

[0055] Fig. 1A and Fig. Figure 1B shows diagrams of microRNA expression released in the extracellular vesicle and intracellularly, obtained by microarray analysis and qPCR analysis, which were used in the sensitization process for chemotherapy drugs according to the preferred embodiment of the present invention. Referring now to Fig. 1A and Fig. 1B shows, for example, that the qPCR analysis shows the highest increases of miR-1976, miR-877-3p and miR-4728-3p in pancreatic cancer cells, with an apparent increase of miR-1976.

[0056] If we now Fig. By applying 1B, for example, the amount of miR-1976 in pancreatic cancer cells can be selectively increased by induction of gemcitabine, which can damage the pancreatic cancer cells.

[0057] Fig. 2A and Fig. Figure 2B is a diagram of the miR-1976-binding target obtained by cloning specific genes, and a diagram of potential target genes (related to cancer cell apoptosis) compared with bioinformatics (XAF1) applied in the chemotherapy drug sensitization method according to the preferred embodiment of the present invention. Referring now to Fig. 2A and Fig. 2B includes, for example, the method for sensitizing to chemotherapy drugs according to the preferred embodiment of the present invention: screening at least one microRNA-binding target within a predetermined cell using a specific gene cloning technology; providing a target-binding RNA fragment to be transcribed or derived into the microRNA-binding target; providing sequential analysis for the target-binding RNA fragment and determining a target gene and its corresponding position using blast analysis to obtain a plurality of miR-1976-binding targets; comparing and searching for a cell apoptosis gene or a related gene with the plurality of miR-1976-binding targets and predicting miR-1976 targets using a target scan tool to obtain an XAF1 gene or a related gene that is a control target of miR-1976.

[0058] Fig. Figures 3A to 3C are diagrams of miR-1976 expression obtained by qPCR analysis and Western blot analysis, which were used in the sensitization process for chemotherapy drugs according to the preferred embodiment of the present invention. Referring now to Fig. 3A to 3C describe the method for sensitizing cancer cells to chemotherapy drugs according to the present invention, for example, by providing a higher level of miR-1976 expression in cancer cells than by simulating cell damage to induce an increase in miR-1976, or by cells affected by the effect of miR-1976-containing extracellular vesicles. Apparently, when cancer cells show a higher level of miR-1976 expression, the level of XAF1 is relatively reduced, and the level of a pancreatic cancer cell apoptosis marker (i.e., cleaved PARP) is also specifically reduced.

[0059] Fig. 4A and Fig. Figure 4B shows diagrams of cell apoptosis of BxPC-3 and AsPC-1 pancreatic cancer cells transfected with AntagomiR-1976, obtained by Western blot analysis performed in the chemotherapy drug sensitization procedure according to the preferred embodiment of the present invention. Referring now to Fig. Figure 4A shows, for example, results from the pancreatic cancer cell BxPC-3, which was used in the sensitization method for chemotherapy drugs according to the present invention. Referring now to Fig. Figure 4B shows, for example, results of the pancreatic cancer cell AsPC-1, which were used in the sensitization method for chemotherapy drugs according to the present invention.

[0060] Still referring to Fig. 4A and Fig. 4B, for example, shows that the amount of a pancreatic cancer cell apoptosis marker (i.e., split PARP) of transfected pancreatic cancer cells appears to be increased because the miR-1976 of the pancreatic cancer cells is blocked.

[0061] The method for sensitization to chemotherapy drugs according to a preferred embodiment of the present invention can, for example, replace an anthagomir-1976 inhibitor with a miR-1976 inhibitor or an equivalent inhibitor.

[0062] The antagomir used here in the preferred embodiment of the present invention, for example, has a specific structure to form a small interfering RNA (siRNA) molecule which has a specific RNA sequence as the specific sequence of “ACAGCAA”.

[0063] For example, the antagomir-1976 used here in the preferred embodiment of the present invention has a 20-nt RNA sequence as the specific sequence of 5'-ACAGCAAGGAGGGCAGGAGG-3'.

[0064] The antagomir-1976 used here in a further preferred embodiment of the present invention, for example, has a key region which is used to inhibit the nucleotide-1976, wherein the key region includes a specific sequence of “ACAGCAA” having a nucleotide length of 17-42 nucleobases.

[0065] The antagomir-1976 used here in a further preferred embodiment of the present invention, for example, has a miR-1976 inhibitor sequence of 5'-(N)xACAGCAA(N)y, where N is the nucleobase A, U, G or C; x is 0 to 5; and y is 10 to 30.

[0066] Fig. Figures 5A to 5D are diagrams of the cancer cell viability of MIA Capa-2, BxPC-3, and AsPC-1 pancreatic cancer cells with siRNA-1976 transfection, which were used in the chemotherapy drug sensitization method according to the preferred embodiment of the present invention. Referring now to Fig. 5A to 5D, the method for sensitizing to chemotherapy drugs according to the present invention must, for example, use at least one active ingredient composition for sensitizing to chemotherapy drugs (e.g. siRNA-1976 or equivalent composition for sensitizing to drugs) and at least one chemotherapy drug, which impairs the viability of pancreatic cancer cells.

[0067] With continuous reference to Fig.5A to 5D, the method for sensitizing to chemotherapy drugs according to the present invention cannot impair the viability of pancreatic cancer cells if only an active ingredient composition for sensitizing to chemotherapy drugs is used in the chemotherapy treatment, without using a chemotherapy drug.

[0068] Although the invention has been described in detail with reference to its currently preferred embodiment, it is understandable to the person skilled in the art that various modifications can be made without departing from the meaning and scope of the invention as set out in the claims in the appendix.

Claims

[1] Methods for sensitizing patients to chemotherapy drugs, comprising: Transfecting an antisense oligonucleotide into a cancer cell to form a transfected cancer cell to block the cancer cell's miR-1976 during future treatment; Delivering a treatment with a cancer drug to the transfected cancer cell whose miR-1976 is blocked by the antisense oligonucleotide; and Offering an increased degree of sensitivity to the cancer drug through the transfected cancer cell during treatment with a cancer drug, as the miR-1976 of the cancer cell is blocked. [2] Method for sensitizing to chemotherapy drugs according to claim 1, wherein the cancer cell is selected from a pancreatic cancer cell. [3] Method for sensitizing to chemotherapy drugs according to claim 1, further comprising: Searching for at least one molecular target in the cancer cell; Screening at least one microRNA-binding target within a predetermined cell; Providing a target-binding RNA fragment to be transcribed or extracted into the microRNA-binding target; Providing sequential analysis for the target-binding RNA fragment and determining a target gene and its corresponding position to obtain a variety of miR-1976-binding targets; and Searching for an XAF1 gene with a multitude of miR-1976 binding targets and providing an antisense oligonucleotide transfection procedure for the cancer cell to block the cancer cell's miR-1976. [4] Method for sensitizing to chemotherapy drugs according to claim 3, wherein the at least one microRNA-binding target is screened by cloning specific genes. [5] Method for sensitizing to chemotherapy drugs according to claim 3, wherein the target gene and its corresponding position are obtained by blast analysis. [6] Method for sensitizing to chemotherapy drugs according to claim 1, wherein expression of miR-1976 is obtained by microarray analysis, qPCR analysis or RNA sequence analysis. [7] Active ingredient composition for sensitization to chemotherapy drugs, comprising: an antisense oligonucleotide provided to treat a cancer cell with an antisense oligonucleotide transfection procedure to create a transfected cancer cell to block the cancer cell's miR-1976 during future treatment; and a cancer drug that is delivered to the transfected cancer cell, which is administered as a combination to increase sensitivity; wherein the cancer drug is provided to treat the transfected cancer cell whose miR-1976 is blocked, the miR-1976-blocked cancer cell offering an increased degree of sensitivity to the cancer drug. [8] Active ingredient composition for sensitization to chemotherapy drugs according to claim 7, wherein the cancer cell is selected from a pancreatic cancer cell. [9] Active ingredient composition for sensitization to chemotherapy drugs according to claim 7, wherein the combination for sensitization causes a reduction of miR-1976 in the extracellular vesicle. [10] Active ingredient composition for sensitization to chemotherapy drugs according to claim 7, wherein the combination for increasing sensitivity causes an increase in cancer cell death or a reduction in cancer cell growth. [11] Active ingredient composition for sensitization to chemotherapy drugs according to claim 7, wherein the combination for increasing sensitivity causes an increase in a cancer cell apoptosis marker. [12] Active ingredient composition for sensitization to chemotherapy drugs according to claim 11, wherein the cancer cell apoptosis marker is selected from cleaved PARP. [13] Active ingredient composition for sensitization to chemotherapy drugs according to claim 11, wherein the cancer cell apoptosis marker corresponds to an XAF1 gene of the cancer cell. [14] Test procedures for sensitization to chemotherapy drugs, including: Transfecting an antisense oligonucleotide into a cancer cell to form a transfected cancer cell to block the cancer cell's miR-1976 during future treatment; Delivering a treatment with a cancer drug to the transfected cancer cell whose miR-1976 is blocked by the antisense oligonucleotide; Offering an increased degree of sensitivity to the cancer drug by the transfected cancer cell during treatment with a cancer drug, since the cancer cell's miR-1976 is blocked; and Testing a quantity of miR-1976 in a blood sample after completion of treatment with a cancer drug. [15] Test method for sensitization to chemotherapy drugs according to claim 14, wherein the cancer cell is selected from a pancreatic cancer cell.

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