Use of Rv0927c protein in the preparation of anti-tumor drugs

Abstract

The present invention relates to the field of medicine, and in particular to the use of Rv0927c protein in the preparation of anti-tumor drugs, wherein the Rv0927c protein includes an amino acid sequence as shown in SEQ ID NO: 1. The Rv0927c protein may be a natural Rv0927c protein or a recombinant Rv0927c protein. The Rv0927c protein is a protein having an amino acid sequence as shown in SEQ ID NO: 1, or a fusion protein formed by connecting a protein having an amino acid sequence as shown in SEQ ID NO: 1 with a carrier protein, an antibody or other amino acid fragments of any length. The tumor is selected from lung cancer. The Rv0927c protein does not produce excessive immune rejection reactions, does not damage the host's immune defense ability while inhibiting the proliferation of lung cancer cells, and can avoid causing side effects.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to the use of Rv0927c protein in the preparation of anti-tumor drugs. Background Art

[0002] Lung cancer is one of the most common malignant tumors in the world today. Its morbidity and mortality rates are increasing year by year, and its mortality rate is also among the highest among all malignant tumors. Lung cancer can be divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) according to histopathology. The most common non-small cell lung cancer is lung adenocarcinoma. Due to its invasive biological behavior and lack of specific symptoms and signs in the early stage, most patients are already in the advanced stage when diagnosed and cannot undergo surgical resection, resulting in a 5-year survival rate of only about 15% for lung adenocarcinoma.

[0003] Chemotherapy is currently an effective measure for the treatment of lung cancer. With the development of molecular biotechnology, new molecular classifications of lung cancer are constantly being discovered. Designing targeted drugs based on lung adenocarcinoma driver proteins and conducting personalized treatments have been successful. However, almost all lung adenocarcinoma patients will develop acquired drug resistance during treatment. Therefore, finding new inhibitors for lung adenocarcinoma cells is of great significance for improving the current status of lung adenocarcinoma cell resistance and developing new and effective lung adenocarcinoma treatments. Therefore, there is an urgent need in this field to develop a drug that can effectively inhibit the activity of lung adenocarcinoma cells. Summary of the invention

[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a use of Rv0927c protein in the preparation of anti-tumor drugs to solve the problems in the prior art.

[0005] To achieve the above-mentioned object and other related objects, the present invention provides Rv0927c protein, polynucleotides encoding Rv0927c protein, expression vectors, viruses or host cells containing polynucleotides encoding Rv0927c protein for use in preparing anti-tumor drugs, wherein the Rv0927c protein comprises an amino acid sequence as shown in SEQ ID NO:1.

[0006] The Rv0927c protein may be a natural Rv0927c protein or a recombinant Rv0927c protein.

[0007] The Rv0927c protein can be a protein with an amino acid sequence as shown in SEQ ID NO: 1, or a fusion protein formed by connecting the protein with an amino acid sequence as shown in SEQ ID NO: 1 or a similar protein thereof with a carrier protein, an antibody or other amino acid fragments of any length.

[0008] The Rv0927c protein can also be a modified protein produced by modifying the amino acid sequence of the protein shown in SEQ ID NO: 1 or any sequence of similar proteins.

[0009] The polynucleotide may be in the form of DNA or RNA.

[0010] The expression vector is a prokaryotic expression vector or a eukaryotic expression vector.

[0011] The host cell can be a prokaryotic cell or a eukaryotic cell.

[0012] The tumor is selected from lung cancer. The lung cancer is selected from small cell lung cancer or non-small cell lung cancer.

[0013] The present invention also provides a retrovirus, wherein the retrovirus contains a polynucleotide encoding the Rv0927c protein, and the Rv0927c protein includes the amino acid sequence shown in SEQ ID NO:1.

[0014] The present invention also provides the use of Rv0927c protein, polynucleotides encoding Rv0927c protein, expression vectors, viruses or host cells containing polynucleotides encoding Rv0927c protein in preparing products for inhibiting tumor cell proliferation or promoting tumor cell apoptosis, wherein the Rv0927c protein includes the amino acid sequence shown in SEQ ID NO: 1.

[0015] As described above, the use of the Rv0927c protein of the present invention in the preparation of anti-tumor drugs has the following beneficial effects: the small molecule protein Rv0927c of the present invention will not produce excessive immune rejection reaction; secondly, Rv0927c has a dose-dependent effect on inhibiting the proliferation of lung cancer cells, and can produce moderate inhibition by reducing the concentration without damaging the host's immune defense ability, thereby avoiding side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Shows the expression and identification of the active protein of Rv0927c expressed in Escherichia coli prokaryotically.

[0017] Figure 2 It was shown that Rv0927c recombinant protein inhibited the proliferation level of human lung cancer A549 cells in vitro.

[0018] Figure 3 It was shown that Rv0927c recombinant protein promoted the apoptosis level of human lung cancer cells A549.

[0019] Figure 4Showing the expression identification of Rv0927c active protein expressed by retroviral vector, 1: pMSCVpuro empty vector group; 2: pMSCVpuro-His-Rv0927c recombinant vector group.

[0020] Figure 5 It was shown that retroviral vector expressing Rv0927c inhibited the growth of lung cancer A549 cells in vivo. DETAILED DESCRIPTION

[0021] The present invention provides Rv0927c protein, a polynucleotide encoding the Rv0927c protein, an expression vector, a virus or a host cell containing the polynucleotide encoding the Rv0927c protein for use in preparing an anti-tumor drug, wherein the Rv0927c protein comprises an amino acid sequence as shown in SEQ ID NO: 1 or a sequence having a similarity of more than 90%, more than 95%, more than 97%, more than 98% or more than 99% to the sequence as shown in SEQ ID NO: 1:

[0022] MILDMFRLDDKVAVITGGGRGLGAAIALAFAQAGADVLIASRTSSELDAVAEQIRAAGRRAHTVAADLAHPEVTAQLAGQAVGAFGKLDIVVNNVGGTMPNTLLSTSTKDLADAFAFNVGTAHALTVAAVPLM LEHSGGGSVINISSTMGRLAARGFAAYGTAKAALAHYTRLAALDLCPRVRVNAIAPGSILTSALEVVAANDELRAPMEQATPLRRLGDPVDIAAAAVYLASPAGSFLTGKTLEVDGGLTFPNLDLPIPDL(SEQ ID NO:1)

[0023] Proteins comprising sequences having a similarity of 90% or more, 95% or more, 97% or more, 98% or more, or 99% or more to the sequence shown in SEQ ID NO: 1 are referred to as similar proteins to Rv0927c.

[0024] The Rv0927c protein may be a natural Rv0927c protein or a recombinant Rv0927c protein.

[0025] Furthermore, the Rv0927c protein includes a protein formed by any Rv0927c protein sequence derived from humans, mice, dogs, cattle, pigs, etc.

[0026] The Rv0927c protein can be a protein with an amino acid sequence as shown in SEQ ID NO: 1, or a fusion protein formed by connecting the protein with an amino acid sequence as shown in SEQ ID NO: 1 or a similar protein thereof with a carrier protein, an antibody or other amino acid fragments of any length.

[0027] Furthermore, the Rv0927c protein can also be a modified protein produced by modifying the amino acid sequence of the protein shown in SEQ ID NO: 1 or any sequence of similar proteins.

[0028] Furthermore, the modification method of the protein modifier is selected from glycosylation modification, fatty acid modification, acylation modification, Fc fragment fusion, albumin fusion, polyethylene glycol modification, dextran modification, heparin modification, polyvinyl pyrrolidone modification, polyamino acid modification, polysialic acid modification, chitosan and its derivatives modification, lectin modification, sodium alginate modification, carbomer modification, polyvinyl pyrrolidone modification, hydroxypropyl methylcellulose modification, hydroxypropyl cellulose modification, acetylation modification, formylation modification, phosphorylation modification, methylation modification or sulfonation modification and one or more of other pharmaceutically available polypeptide / protein drug modification methods.

[0029] The similar proteins, fusion proteins or protein modifications of the Rv0927c protein all have the functions of the Rv0927c protein and can bind to the ligand of Rv0927c.

[0030] The polynucleotide may be in the form of DNA or RNA. The DNA may be selected from cDNA, genomic DNA or artificially synthesized DNA. The DNA may be single-stranded or double-stranded.

[0031] In certain embodiments of the invention, the polynucleotide sequence such as SEQ ID NO:2 shows: ATGATCCTGGATATGTTCCGTCTTGACGACAAGGTTGCCGTCATCACCGGCGGCGGCCGCGGCCTGGGAGCAGCCATCGCATTGGCTTTCGCCCAGGCCGGCCGGATGTCCTCATCGCTTCACGAACATCATCCGAGCTTGACGCTGTCGCCGAACAGATCCGCGCTGCCGGCCGCCGCGCCCACACCGTT GCCGCCGATCTGGCCCATCCCGAGGTGACCGCGCAGCTGGCTGGTCAGGCCGTCGGAGCTTTCGGGAAGCTCGACATCGTCGTCAACAACGTTGGCGGCACCATGCCCAACACGCTGCTAAGCACCTCGACCAAGGACCTCGCGGACGCCTTCGCCTTCAACGTGGGCACCGCCCACGCGCTGACCGTCGCGGCGGTGCC GTTGATGCTGGAACACTCCGGCGGCGGCAGCGTGATCAACATCAGTCCCACCATGGGCCGGCTGGCGGCGCGGGGTTTCGCCGCCTACGGCACCGCCAAGGCCGCACTGGCCCACTACACCCGGCTGGCGGCGCTGGACCTGTGCCCACGCGTCCGGGTCAATGCGATCGCGCCGGGTTCCATCCTGACCTCGGCGCTGG AGGTGGTAGCCGCCAACGACGAGCTGCGTGCGCCGATGGAGCAGGCGACACCACTGCGCCGGCTCGGTGACCCCGTCGACATTGCAGCTGCGGCAGTGTATTTGGCCTCTCCGGCGGGCAGCTTCCTGACCGGCAAGACGTTGGAGGTCGACGGCGGACTCACCTTCCCCAATCTCGACCTTCCCATTCCGGACCTGTGA

[0032] Those skilled in the art are familiar with methods that can be used to construct the expression vector. These methods include, for example, recombinant DNA technology, DNA synthesis technology, etc. The DNA encoding the protein can be effectively connected to an appropriate promoter in the expression vector to guide mRNA synthesis and then express the protein.

[0033] Preferably, the expression vector is a prokaryotic expression vector or a eukaryotic expression vector.

[0034] The prokaryotic expression vector is selected from an Escherichia coli expression vector, a Bacillus subtilis expression vector or a Streptomyces expression vector. In a preferred embodiment, the prokaryotic expression vector is selected from an Escherichia coli expression vector. Compared with other expression systems, the Escherichia coli expression system has a clear genetic background, a short culture cycle, a high expression level of the target gene, and strong anti-pollution ability. The Escherichia coli expression vector, such as pET32a, can be stably expressed in Escherichia coli.

[0035] The eukaryotic expression vector is selected from a yeast expression vector, an insect expression vector or a mammalian expression vector. The mammalian expression vector is selected from a retroviral expression vector, a lentiviral expression vector, an adenoviral expression vector, and an adeno-associated viral expression vector. In a preferred embodiment, the eukaryotic expression vector is selected from a retroviral expression vector, which can be stably expressed in a cell line, and the retroviral vector is, for example, pMSCV.

[0036] The virus is selected from a lentivirus, an adenovirus, an adeno-associated virus or a retrovirus.

[0037] The host cell may be a prokaryotic cell, such as a bacterial cell, or a eukaryotic cell. Representative examples of the host cell include Escherichia coli, 293T cells, or human lung cancer cells A549.

[0038] The Rv0927c protein can be prepared by existing techniques, such as chemical synthesis, or by culturing host cells under conditions suitable for expressing the protein and isolating the protein from the culture. The method for isolating the protein from the culture can be extracted by nickel column purification.

[0039] In the present invention, the tumor is, for example, lung cancer, renal cell carcinoma, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric cancer, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic cancer; leukemia, lymphoma, myeloma, mycoses fungoids, Merkel cell carcinoma and other malignant blood diseases, such as classical Hodgkin lymphoma (CHL), primary mediastinal large B-cell lymphoma, T cell / histiocyte-rich B-cell lymphoma, EBV-positive and negative PTLD and EBV-associated diffuse large B-cell lymphoma (DLBCL), plasmablastic lymphoma, extranodal NK / T cell lymphoma, nasopharyngeal carcinoma and HHV8-associated primary effusion lymphoma.

[0040] In a preferred embodiment, the tumor is selected from lung cancer, and the lung cancer is selected from small cell lung cancer or non-small cell lung cancer.

[0041] The anti-tumor drug achieves the anti-tumor effect by inhibiting the proliferation of tumor cells or promoting the apoptosis of tumor cells.

[0042] The present invention also provides a retrovirus, wherein the retrovirus contains a polynucleotide encoding the Rv0927c protein, and the Rv0927c protein includes the amino acid sequence shown in SEQ ID NO:1.

[0043] The present invention also provides a method for treating tumors, which comprises administering to a subject a therapeutically effective amount of any one or more of the following: Rv0927c protein, a polynucleotide encoding the Rv0927c protein, an expression vector containing the polynucleotide encoding the Rv0927c protein, or a host cell.

[0044] "Subjects" include, but are not limited to, animals, preferably mammals; the mammals are preferably rodents, artiodactyls, perissodactyls, lagomorphs, primates, etc. The mammals include, for example, humans, non-human primates (e.g., monkeys), mice, pigs, cows, goats, rabbits, rats, guinea pigs, hamsters, horses, monkeys, sheep, or other non-human mammals; non-mammals include, for example, non-mammalian vertebrates, such as birds (e.g., chickens or ducks) or fish, and non-mammalian invertebrates. The subject can be a human, such as a patient with a low immune function or cancer.

[0045] "Tumor" in this application refers to any medical condition mediated by tumor or malignant cell growth, proliferation or metastasis, and causes solid tumors and non-solid tumors such as leukemia. "Tumor" in the present invention refers to the solid material of tumor and / or malignant cells.

[0046] The "therapeutically effective amount" or "effective dose" in the present invention refers to the dose or concentration of a drug that is effective in treating a related disease or condition. For example, for the protein disclosed in the present invention, the therapeutically effective amount is the dose or concentration at which the protein can eliminate all or part of a tumor, inhibit or slow tumor growth, inhibit the growth or reproduction of cells that mediate a cancerous state, inhibit tumor cell metastasis, alleviate any symptoms or markers associated with a tumor or cancerous state, prevent or delay the development of a tumor or cancerous state, or some combination of the above.

[0047] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0048] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0049] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0050] Example 1 Recombinant Rv0927c protein inhibits proliferation of human lung cancer cells A549 in vitro

[0051] (1) Prokaryotic expression and identification of recombinant Rv0927c protein

[0052] First, design primers with EcoRI and Hind III double restriction sites:

[0053] Forward Primer:

[0054] 5'-GCTGATATCGGATCCGAATTCATGATCCTGGATATGTTCCGTCTT-3' (SEQ ID NO: 3)

[0055] Reverse Primer:

[0056] 5'-CTCGAGTGCGGCCGCAAGCTTTCACAGGTCCGGAATGGGA-3' (SEQ ID NO: 4)

[0057] Then, the genome was extracted from Mycobacterium tuberculosis (purchased from ATCC) as a template, and the sequences of SEQ ID NO: 3 and SEQ ID NO: 4 were used as amplification primers to PCR amplify the Rv0927c (sequence as shown in SEQ ID NO: 2) gene, and then agarose gel electrophoresis was performed to recover the Rv0927c product, and the Rv0927c product was connected to the pET32a plasmid (purchased from Novagen), and then DH5α (purchased from TAKAR) was transformed, and the resistant LB solid medium was coated for screening. Positive clones were picked for identification, and the prokaryotic expression plasmid pET32a-Rv0927c was obtained after sequencing (Nanjing GenScript Biotechnology Co., Ltd.) was correct. The confirmation results showed that the Rv0927c gene expression sequence in the prokaryotic expression plasmid pET32a-Rv0927c was as shown in SEQ ID NO: 2.

[0058] Pick a positive single colony, place it in 4mL of ampicillin-resistant LB liquid medium, and culture it in a 37°C constant temperature shaker overnight. The next day, the recombinant bacteria were inoculated into ampicillin-resistant LB liquid medium at a ratio of 1:100, cultured in a 37°C constant temperature shaker with an OD value between 0.4-0.6, and IPTG was added to a final concentration of 0.5mmol / L, and induced in a constant temperature shaker at 37°C 150rpm for 6h. The bacteria were collected by centrifugation and washed three times with sterile PBS. The bacteria were lysed using an ultrasonic lysator in an ice-water mixture (30W, lysis 3s, interval 5s). The supernatant was then collected by centrifugation at 10000rpm for 10min. The supernatant after centrifugation was removed and the precipitate was resuspended with an equal volume of sterile PBS. The supernatant collected after centrifugation was purified using a Ni-Agarose His tag protein purification kit. First, the column was pretreated, and 7.5 mL of sterile ultrapure water, 12.5 mL of Charge buffer, and 7.5 mL of Binding buffer were added to rinse the purification column with the His tag, and then the sample was added to the column, and finally 25 mL of Binding buffer, 15 mL of Washing buffer, and 15 mL of Elute buffer were added. The column was covered with a stopper and allowed to stand at room temperature for 20 min. The purified protein was collected in a sterile 1.5 mL finger tube and analyzed by SDS-PAGE and Western Blot, respectively ( Figure 1 ). In addition, sequencing was performed to confirm that the amino acid sequence of the expressed protein was correct, as specifically shown in SEQ ID NO: 1.

[0059] (2) Recombinant Rv0927c protein inhibits proliferation of human lung cancer A549 cells in vitro

[0060] A549 cells were cultured at 5 × 10 4The cells / well were plated into 24-well plates. After overnight, the cells were stimulated with different concentrations of recombinant Rv0927c protein. At 0h, 24h, 48h, and 72h, the cell supernatant was discarded and CCK-8 diluent (100μL complete DMEM + 10μL CCK-8) was added. The cells were cultured at 37℃ for 1h and the OD value was measured by microplate reader. 450 The results showed that Rv0927c protein significantly inhibited the proliferation activity of A549 cells in vitro ( Figure 2 ).

[0061] Example 2 Recombinant Rv0927c protein induces apoptosis level of human lung cancer cell A549

[0062] A549 cells were cultured at 1×10 5 Cells / well were plated into 24-well plates. When the cells grew to 70%, 10 μg / mL recombinant Rv0927c protein was used to stimulate the cells. After 24 hours, the cell supernatant was collected, the cell plate was washed once with PBS, the cells were digested with EDTA-free trypsin, and PBS was added to resuspend the cells. At the same time, the collected cell supernatant was centrifuged and the pellet was resuspended with the cell suspension. The cells in the cell suspension were counted and the concentration was adjusted to 1×10 6 cells / mL, washed twice by centrifugation with 1×Binding Buffer. Resuspend the cells with 100μL1×Binding Buffer, add 10μL of Annexin V-FITC, mix well, and incubate at room temperature in the dark for 15min. Wash twice by centrifugation with 1×Binding Buffer, resuspend the cells with 500μL 1×Binding Buffer, place on ice, add 5μL of PI solution 2min before detection, and detect using flow cytometer. The results showed that the proportion of apoptotic cells in the untreated group was about 11.8%, while the proportion of apoptotic cells in the Rv0927c recombinant protein stimulation group reached 35.2%. Statistical analysis results showed that Rv0927c protein significantly promoted the apoptosis level of A549 cells ( Figure 3 ).

[0063] Example 3 Rv0927c retrovirus inhibits the growth of lung cancer A549 cells in vivo

[0064] ⑴ Construction and identification of recombinant retroviral plasmid pMSCV-Rv0927c

[0065] First, design primers and add a His tag sequence at the C-terminus:

[0066] Forward Primer:

[0067] 5'-GCCGGAATTAGATCTCTCGAGATGATCCTGGATATGTTCCGTCTT-3' (SEQ ID NO: 5)

[0068] Reverse Primer:

[0069] 5'-CTCCCCTACCCGGTAGAATTCTCAGTGATGATGGTGATGATGCAGGTCCGGAATGGGA-3'(SEQID NO: 6)

[0070] The target gene was amplified and recovered using a high-fidelity enzyme, and then ligated to the pMSCV vector (Clontech, Cat. No. 634401) using a one-step ligase, and heat-transferred to DH5α. Single clones were picked for sequencing and identification. HEK293 cells were cultured at 2×10 6 The cells / dish were spread into a 10 cm culture dish. After 16 hours, pMSCV-Rv0927c and pcl-Ampho were transfected into the cells using Lipofectamine 3000. Lipofectamine premix: 250 μL Opti-MEM + 9 μL 3000; Prepare DNA premix: 250μL Opti-MEM+pMSCV-Rv0927c(4μg)+pcl-Ampho(2μg). Slowly drop the DNA premix into the diluted 3000 reagent, incubate at room temperature for 5-10 minutes, slowly drip the complex into the plate, and culture at 37℃ for 24h. Change the medium for the transfected cells and continue to culture at 37℃. Collect the cell supernatant at 48h and 72h, centrifuge to remove cell debris, and then filter with a 0.45μm filter, mark as virus stock solution, and store in a -80℃ refrigerator.

[0071] ⑵ Retroviral expression of Rv0927c inhibits the growth of lung cancer A549 cells in vivo

[0072] A549 cells were cultured at 1×10 5 Cells / well were plated into 6-well plates. After 16 hours, the supernatant was discarded and the virus premix was added: 1mL complete DMEM medium + 1mL retrovirus + polybrene with a final concentration of 10μg / mL. The cells were centrifuged and infected at room temperature for 90 minutes. After the infection, fresher medium was added and cultured at 37°C. After 24 hours, the complete DMEM medium containing puromycin was replaced (the concentration of puromycin was the minimum concentration that killed all normal A549 cells). After the cells were grown, they were collected and identified by Western blot ( Figure 4 ), indicating that Rv0927c protein was successfully and stably expressed in A549 cells.

[0073] The retrovirus-infected cells were adjusted to 6 × 10 7 The cells were injected into the subcutaneous tissue of mice, and the tumor formation of mice was observed every 3 days. The tumors formed were removed after 45 days. The results showed that the volume and size of the tumors formed in the Rv0927c expression cell injection group were significantly reduced, indicating that Rv0927c expression inhibited the growth of mouse tumors ( Figure 5 ).

[0074] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and the variations of the methods in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of the present invention. Sequence Listing <110> Yangzhou University <120> Use of Rv0927c protein in the preparation of anti-tumor drugs <160> 6 <170> SIPOSequenceListing 1.0 <210> 1 <211> 263 <212> PRT <213> Artificial Sequence <400> 1 Met Ile Leu Asp Met Phe Arg Leu Asp Asp Lys Val Ala Val Ile Thr 1 5 10 15 Gly Gly Gly Arg Gly Leu Gly Ala Ala Ile Ala Leu Ala Phe Ala Gln 20 25 30 Ala Gly Ala Asp Val Leu Ile Ala Ser Arg Thr Ser Ser Ser Glu Leu Asp 35 40 45 Ala Val Ala Glu Gln Ile Arg Ala Ala Gly Arg Arg Ala His Thr Val 50 55 60 Ala Ala Asp Leu Ala His Pro Glu Val Thr Ala Gln Leu Ala Gly Gln 65 70 75 80 Ala Val Gly Ala Phe Gly Lys Leu Asp Ile Val Val Asn Asn Val Gly 85 90 95 Gly Thr Met Pro Asn Thr Leu Leu Ser Thr Ser Thr Lys Asp Leu Ala 100 105 110 Asp Ala Phe Ala Phe Asn Val Gly Thr Ala His Ala Leu Thr Val Ala 115 120 125 Ala Val Pro Leu Met Leu Glu His Ser Gly Gly Gly Ser Val Ile Asn 130 135 140 Ile Ser Ser Thr Met Gly Arg Leu Ala Ala Arg Gly Phe Ala Ala Tyr 145 150 155 160 Gly Thr Ala Lys Ala Ala Leu Ala His Tyr Thr Arg Leu Ala Ala Leu 165 170 175 Asp Leu Cys Pro Arg Val Arg Val Asn Ala Ile Ala Pro Gly Ser Ile 180 185 190 Leu Thr Ser Ala Leu Glu Val Val Ala Ala Asn Asp Glu Leu Arg Ala 195 200 205 Pro Met Glu Gln Ala Thr Pro Leu Arg Arg Leu Gly Asp Pro Val Asp 210 215 220 Ile Ala Ala Ala Ala Val Tyr Leu Ala Ser Pro Ala Gly Ser Phe Leu 225 230 235 240 Thr Gly Lys Thr Leu Glu Val Asp Gly Gly Leu Thr Phe Pro Asn Leu 245 250 255 Asp Leu Pro Ile Pro Asp Leu 260 <210> 2 <211> 792 <212> DNA <213> Artificial Sequence <400> 2 atgatcctgg atatgttccg tcttgacgac aaggttgccg tcatcaccgg cggcggccgc 60 ggcctgggag cagccatcgc attggctttc gcccaggccg gcgcggatgt cctcatcgct 120 tcacgaacat catccgagct tgacgctgtc gccgaacaga tccgcgctgc cggccgccgc 180 gcccacaccg ttgccgccga tctggcccat cccgaggtga ccgcgcagct ggctggtcag 240 gccgtcggag ctttcgggaa gctcgacatc gtcgtcaaca acgttggcgg caccatgccc 300 aacacgctgc taagcacctc gaccaaggac ctcgcggacg ccttcgcctt caacgtgggc 360 accgcccacg cgctgaccgt cgcggcggtg ccgttgatgc tggaacactc cggcggcggc 420 agcgtgatca acatcagctc caccatgggc cggctggcgg cgcggggttt cgccgcctac 480 ggcaccgcca aggccgcact ggcccactac acccggctgg cggcgctgga cctgtgccca 540 cgcgtccggg tcaatgcgat cgcgccgggt tccatcctga cctcggcgct ggaggtggta 600 gccgccaacg acgagctgcg tgcgccgatg gagcaggcga caccactgcg ccggctcggt 660 gaccccgtcg acattgcagc tgcggcagtg tatttggcct ctccggcggg cagcttcctg 720 accggcaaga cgttggaggt cgacggcgga ctcaccttcc ccaatctcga ccttcccatt 780 ccggacctgt ga 792 <210> 3 <211> 45 <212> DNA <213> Artificial Sequence <400> 3 gctgatatcg gatccgaatt catgatcctg gatatgttcc gtctt 45 <210> 4 <211> 40 <212> DNA <213> Artificial Sequence <400> 4 ctcgagtgcg gccgcaagct ttcacaggtc cggaatggga 40 <210> 5 <211> 45 <212> DNA <213> Artificial Sequence <400> 5 gccggaatta gatctctcga gatgatcctg gatatgttcc gtctt <210> 6 <211> 58 <212> DNA <213> Artificial Sequence <400> 6 ctcccctacc cggtagaatt ctcagtgatg atggtgatga tgcaggtccg gaatggga

Claims

1. Use of Rv0927c protein, a polynucleotide encoding Rv0927c protein, an expression vector containing the polynucleotide encoding Rv0927c protein, a virus containing the polynucleotide encoding Rv0927c protein, or a host cell containing the polynucleotide encoding Rv0927c protein in the preparation of an anti-tumor drug, wherein the amino acid sequence of the Rv0927c protein is as shown in SEQ ID NO: 1, and the anti-tumor drug is a drug for non-small cell lung cancer.

2. The use according to claim 1, characterized in that The polynucleotide is in the form of DNA or RNA.

3. The use according to claim 1, characterized in that The sequence of the polynucleotide is shown in SEQ ID NO:

2.

4. The use according to claim 1, characterized in that The expression vector is a prokaryotic expression vector or a eukaryotic expression vector.

5. The use according to claim 4, characterized in that The eukaryotic expression vector is selected from a yeast expression vector, an insect expression vector or a mammalian expression vector.

6. The use according to claim 5, characterized in that The mammalian expression vector is selected from a retrovirus expression vector, an adenovirus expression vector or an adeno-associated virus expression vector.

7. The use according to claim 1, characterized in that The virus is selected from adenovirus, adeno-associated virus or retrovirus, and / or the host cell is a prokaryotic cell or a eukaryotic cell.

8. The use according to claim 1, characterized in that The anti-tumor drug is anti-tumor by inhibiting tumor cell proliferation or promoting tumor cell apoptosis.

9. A retrovirus, characterized in that The retrovirus contains a polynucleotide encoding the Rv0927c protein, and the amino acid sequence of the Rv0927c protein is shown in SEQ ID NO:

1.

10. Use of Rv0927c protein, a polynucleotide encoding Rv0927c protein, an expression vector containing a polynucleotide encoding Rv0927c protein, a virus containing a polynucleotide encoding Rv0927c protein, or a host cell containing a polynucleotide encoding Rv0927c protein in the preparation of a product for inhibiting tumor cell proliferation or promoting tumor cell apoptosis, wherein the amino acid sequence of the Rv0927c protein is as shown in SEQ ID NO: 1, and the tumor is non-small cell lung cancer.

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