Application of BTF3L4 gene and its RNAi interference system

Abstract

The present invention discloses the application of the BTF3L4 gene and its RNAi interference system, which relates to the field of biomedical technology. The BTF3L4 gene and the RNAi interference system of the BTF3L4 gene of the present invention are used for preparing drugs for treating glioma. The RNAi interference system can efficiently knockdown the BTF3L4 gene / protein in glioma cells, inhibit the invasion and migration of cancer cells, and has simple operation and high efficiency. The present invention also discloses using the BTF3L4 gene / protein as a target for precision therapy to prepare a kit for diagnosing or predicting prognosis for glioma and other diseases with high expression of the BTF3L4 gene / protein, and it is also used for preparing targeted drugs.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and more specifically, relates to the application of the BTF3L4 gene (gene ID: 91408 and protein ID: Q96K17) and its RNAi interference system in the preparation of drugs for treating glioma. Background Art

[0002] Glioma is one of the most common malignant brain tumors globally. The mortality rate within one year of diagnosis is nearly 80%. Even after targeted therapy, the prognosis of patients is still not ideal. The annual incidence of glioma in China is 5 - 8 per 100,000 people, and the 5-year survival rate of patients is less than 5%. Its 5-year fatality rate is second only to pancreatic cancer and lung cancer. In recent years, with the continuous increase in the incidence of global malignant tumors, it has brought a serious burden to the social economy.

[0003] Although the progress of molecular pathological diagnosis of glioma has identified multiple molecular pathological targets, the clinical treatment for these molecular mutations still has no significant effect. Currently, it is generally believed that the glioma microenvironment consists of different types of cells, which evade immune system surveillance and targeted therapy through various mechanisms. Recent studies have also shown that the response of glioma to treatment has changed significantly. Abnormal molecular regulation of multiple types causes extensive inter-tumor and intra-tumor heterogeneity in primary gliomas. Therefore, the development of clinical diagnostic and treatment biomarkers is crucial for improving the survival chances of glioma patients. With the application of high-throughput technologies such as second-generation, third-generation sequencing, single-cell sequencing, proteomics, and metabolomics, targeted therapy has become a new method for treating malignant tumors in addition to surgery, radiotherapy, and chemotherapy. Targeted therapy blocks the growth of cancer cells by interfering with specific target molecules required for the occurrence and progression of tumors, with strong specificity, obvious efficacy, and few adverse reactions. However, targeted therapy inevitably requires us to find specific molecular markers as treatment targets. The development of glioma is a process involving multiple genes, and there are numerous potential treatment targets. Currently, there is no glioma diagnostic kit with clear efficacy on the market. Therefore, there is an urgent need to develop detection reagents and targeted drugs for effective targets.

[0004] Basic transcription factor 3-like protein 4 (BTF3L4) is a novel cartilage formation-related protein, initially reported as a transcription factor promoting cartilage formation. Transcription factors (TFs) are a class of protein molecules that specifically bind to specific sequences of genes, thereby regulating the target genes to express at a specific intensity in a specific time and space. As an important part of gene expression regulation, TFs are widely involved in physiological and pathological processes in the body. Transcription factors play a key biological role in diseases such as cancer, so they are considered potential therapeutic targets. It is estimated that there are at least 1600 transcription factors in the human genome, about 19% of which are closely related to various diseases including cancer. Given the direct regulatory effect of TFs on target genes, they have a higher specific disease regulatory ability. Therefore, transcription factors can be used as new therapeutic targets and driving factors for glioma progression. At the same time, BTF3L4 is also an oncogene, which can promote the growth and metastasis of thyroid tumors and colorectal cancers by increasing nuclear migration rate, and can also inhibit the growth of gastric cancer cells. In addition, in the central nervous system, BTF3L4 is a gene necessary for neuron morphology and is involved in the morphological formation of brain malignancies. However, the relationship between BTF3L4 and glioma is currently unclear, and its expression and specific regulatory mechanism in glioma are also unknown. Summary of the Invention

[0005] In view of the above problems existing in the prior art, the technical problem to be solved by the present invention is to provide the application of BTF3L4 gene and its RNAi interference system for preparing drugs for treating glioma.

[0006] To solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0007] Application of BTF3L4 gene in preparing drugs for treating glioma.

[0008] Further, the treatment of glioma is to inhibit cell migration.

[0009] Further, the treatment of glioma is to inhibit cell invasion.

[0010] The biomarker for detecting the expression level of BTF3L4 protein in tumor tissues is BTF3L4 gene.

[0011] The kit for detecting the expression level of BTF3L4 protein in tumor tissues contains BTF3L4 gene.

[0012] Application of the RNAi interference system of BTF3L4 gene in preparing drugs for treating glioma, and the sequence of the RNAi interference system of BTF3L4 gene is as follows:

[0013] BTF3L4-sh1: 5'-CCTGATGTTACAGTTTGGTAGATTT-3',

[0014] BTF3L4-sh2: 5'-CAAACTGGAATAGCTAGCATGTGCT-3'.

[0015] Furthermore, the RNAi interference system of the BTF3L4 gene is to knockdown the BTF3L4 gene in glioma tissue cells.

[0016] Furthermore, the RNAi interference system of the BTF3L4 gene is to inhibit the expression of BTF3L4 protein in glioma tissue cells.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] 1) The RNAi interference of the present invention can efficiently knockdown the BTF3L4 gene / protein in gastric cancer cells and inhibit the invasion and migration of glioma tissue cells. This system is simple to operate and has high efficiency.

[0019] 2) The BTF3L4 gene / protein is a target for precision therapy. Kits for diagnosing or predicting prognosis for gliomas and other diseases with high expression of the BTF3L4 gene / protein are prepared, and are also used to prepare targeted drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a diagram showing the differential expression of BTF3L4 in glioma and control non-tumor brain tissue microarrays by multiplex immunofluorescence quantitative analysis;

[0021] Figure 2 It is a diagram showing the expression of BTF3L4 in glioma and control non-tumor brain tissues detected by immunofluorescence (A, A1 are fluorescence staining diagrams of BTF3L4 in glioma tissues; B, B1 are immunofluorescence staining diagrams of BTF3L4 in non-tumor brain tissues) (scale bar = 50um);

[0022] Figure 3 It is a diagram showing the relationship between the differential expression of BTF3L4 protein and the prognosis of glioma patients;

[0023] Figure 4 It is a diagram showing the expression and quantitative analysis of BTF3L4 in 6 pairs of surgically resected glioma tissues and adjacent tissues by Western Blot (A) and the expression and quantitative analysis of BTF3L4 in U87mg cells, U251 cells, SHG44 cells and T98G cells (B);

[0024] Figure 5 It is a diagram showing the protein expression and quantitative analysis in U251 cells after interfering with the BTF3L4 gene;

[0025] Figure 6 Migration impact diagram of glioma cells after interfering with BTF3L4 gene;

[0026] Figure 7 Invasion impact diagram of glioma cells after interfering with BTF3L4 gene;

[0027] Figure 8 Cell cycle impact diagram of glioma cells after interfering with BTF3L4 gene. Detailed implementation mode

[0028] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described below in conjunction with specific embodiments. In the following embodiments, unless otherwise specified, the technical means used are all conventional means well-known to those skilled in the art.

[0029] Collect glioma tissue samples from Nantong University Affiliated Hospital between March 2012 and March 2018, including 197 cases of fresh frozen glioma tissues and 28 cases of control non-tumor brain tissues. These tissue samples were all fixed with formalin, embedded in paraffin, and graded according to the latest WHO diagnostic criteria. All cases were histologically determined by two pathology experts, and the patients had not received immunotherapy, chemotherapy or radiotherapy before surgery, and the clinical case data were detailed and complete.

[0030] The main reagents used in the following embodiments are:

[0031] Opal 7-color immunohistochemistry kit: Perkin Elmer, USA.

[0032] Rabbit anti-human BTF3L4 monoclonal antibody: Biorbyt, UK.

[0033] Horseradish peroxidase-labeled mouse-rabbit secondary antibody (for immunofluorescence experiment): Perkin Elmer, USA.

[0034] Antibody diluent / blocking solution: AKOYA, USA.

[0035] AR6 repair solution: AKOYA, USA.

[0036] Glioma cell lines were purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

[0037] DMEM medium, fetal bovine serum: Gibco, USA.

[0038] BCA protein assay kit: Beyotime Biotechnology Co., Ltd.

[0039] PVDP membrane: Merk-Millipore, Germany.

[0040] GAPDH antibody: Proteintech, USA

[0041] ECL developing solution: Vazyme

[0042] Cell cryopreservation solution: Xinsaimi

[0043] Complete DMEM culture medium: DMEM and fetal bovine serum were added respectively to prepare a complete medium with a final concentration of 10% FBS and stored at 4°C.

[0044] 1×TBST 1L: Take 2.42 g of Tris, 8.0 g of NaCl, 0.5 mL of Tween-20, mix and dissolve, make up the volume to 1L, and store at room temperature.

[0045] 1×Transfer membrane Buffer 1L: 14.4 g of glycine, 3.03 g of Tris, add an appropriate amount of double-distilled water and stir to dissolve, then add 200 mL of anhydrous methanol, make up the volume to 1L, and mix evenly (prepared before use).

[0046] Blocking solution 100 mL: Take 5 g of skim milk powder, add 100 mL of 1×TBS, mix and dissolve (prepared before use).

[0047] Washing solution PBST: 0.8 g of NaCl, 0.02 g of KH2PO4, 0.29 g of Na2HPO4·12H2O, 0.02 g of KCl, 0.05 mL of Tween20, 0.01 g of sodium azide, add double-distilled water to 100 mL, and adjust to pH 7.4.

[0048] Protein lysis reagent: Add 10 μL of protease inhibitor cocktail, 5 μL of PMSF and 5 μL of phosphatase inhibitor to 1 mL of lysis solution.

[0049] The main instruments used in the following examples are as follows:

[0050] Tissue microarray maker: Quick Ray (UNITMA), South Korea; Inverted microscope: Olympus, Japan; Gel imaging system: Tianneng, China; Multifunctional microplate reader: Thermo, USA; Multispectral pathology scanning system: Perkin Elmer, USA.

[0051] Example 1

[0052] 1. Fabricate tissue microarray

[0053] 1) Pathological tissue section

[0054] Take a fresh tissue block (0.5 cm thick) removed by surgery, place it in a pre-prepared 10% formalin solution for fixation, and then dehydrate it with alcohol of gradient concentration until it becomes transparent in xylene; place the transparent tissue block in melted paraffin, and embed it after the paraffin has completely penetrated the tissue block; after cooling and solidifying, make serial sections (5-8 μm thick), and then dry them in an incubator at 45°C.

[0055] 2) HE staining

[0056] Place the hydrated sections in hematoxylin aqueous solution for staining for several minutes; differentiate with hydrochloric acid alcohol and ammonia water; after rinsing with running water for 1 hour, place them in distilled water; then, dehydrate them in 70%, 75%, and 90% alcohol for 5 minutes each; place them in eosin staining solution for staining for 2-3 minutes; after staining, dehydrate the sections with gradient alcohol, then make the sections transparent with xylene, drop neutral gum, cover with a coverslip for sealing, and observe under a microscope to determine the tumor area, and mark the representative cancer nest area on the donor pathological tissue wax block.

[0057] 3) Preparation of glioma tissue microarray

[0058] Mix paraffin and beeswax in a ratio of 1:1 to make a blank recipient wax block; design a 10×7 hole on the wax block, with a total of 350 tissue arrays, and then use a tissue microarray instrument to make a TMA blank wax block; select the most representative tumor area on the marked donor wax block, take a tissue block with a diameter of 2 mm, and take 1 core for each case; transfer the taken tissue cores to the holes of the recipient wax block, and take the corresponding non-tumor brain tissue as a control; heat and fuse the tissue array block in a constant temperature oven at 55°C for 10 minutes, and let it cool to room temperature before it melts quickly to make the recipient wax block integrate with the donor tissue; freeze the tissue microarray at 4°C for about 4 hours, and then use a fully automatic tissue slicer to correct the tissue array block at a speed of 20 mm / revolution until all tissue cores are completely exposed; use the slicer to slice the tissue array block, float the serial sections in cold water to make them unfold naturally, and then transfer the sections to warm water at 45°C for spreading for about 2 minutes. After spreading, stick them on a glass slide treated with anti-falling film and dry them; bake the sections in an environment at 60°C for 3 minutes and continue baking at 58°C for 16 h; store the prepared tissue microarray in a slide box and store it in a refrigerator at -20°C for standby.

[0059] 2. Immunofluorescence staining

[0060] Place the cut paraffin tissue microarray on a baking instrument and bake at 70°C for 1 hour and then at 60°C for 1 hour; Immerse the dried tissue microarray in xylene for 5 minutes and repeat 2 times; After taking it out, perform gradient alcohol dehydration: soak in 100% ethanol for 5 minutes, 95% ethanol for 5 minutes, 75% ethanol for 5 minutes, and finally rinse the tissue microarray with distilled water; Place the tissue microarray on a high-temperature resistant section rack and place it in AR6 repair solution with a pH of 6.0, heat at 100% power for 2.5 minutes, and then heat at 20% power for 15 minutes for high-temperature antigen repair; After naturally cooling to room temperature, take out the microarray in distilled water and rinse it 3 times with PBS for 2 minutes each time; Use an immunohistochemistry pen to draw the approximate tissue range on the tissue microarray, then add 200 μL of primary antibody blocking solution and block for 10 minutes; Add 200 μL of rabbit anti-human BTF3L4 monoclonal antibody working solution (dilution ratio 1:100) to the tissue microarray and incubate overnight at 4°C; The next day, take out the tissue microarray, rewarm for half an hour, recover the primary antibody and then rinse with PBS for 2 minutes and repeat 3 times, then take it out and drain; Add 200 μL of secondary antibody working solution to the tissue microarray and incubate at room temperature for 10 minutes, then rinse with PBS for 2 minutes and repeat 3 times, then take it out and drain; Prepare the fluorescent dye with the required wavelength, add the prepared fluorescent dye to the tissue microarray, incubate in the dark at room temperature for 10 minutes, then rinse with PBS for 2 minutes and repeat 3 times; After the section is dried and made transparent, mount with DAPI.

[0061] Observe the results of immunofluorescence staining under a microscope, and the appearance of staining in the corresponding part of the cells is regarded as a positive manifestation. Use Vectra 3 imaging software to capture each sample at a magnification of 20 times. Use inForm 26.1.0 (Perkin Elmer) to analyze and score the images, and set the threshold for positive or negative cells for each cell. Calculate the percentage of cells in each area and score (0 - 100). The final staining score of BTF3L4 is the product of the staining intensity and the stained area of positive cells. The cut-off point of the BTF3L4 expression score is obtained by X-tile software according to the survival time and survival status. The scoring is as follows: 0 - 50 is low expression or no expression, and 51 - 100 is high expression. All data are processed using statistical software SPSS V.25.0. Measurement data are expressed as mean ± standard deviation. One-way ANOVA is used for inter-group comparison. The Kaplan-Meier survival analysis is used to analyze the relationship between BTF3L4 expression and the prognosis of glioma patients. All test results with P < 0.05 are considered statistically significant. The results are as Figure 1-3 shown. The expression of BTF3L4 in glioma tissues is slightly lower than that in the control non-tumor brain tissues. In tumor patients, patients with high expression of BTF3L4 have a higher glioma grade, a higher degree of malignancy in histopathological grading, a shorter survival period of patients, and a poor prognosis.

[0062] Example 2

[0063] 1. shRNA Design

[0064] First, use Invitrogen Block-iT RNAi Designer to design sequences targeting the BTF3L4 gene, and commission Beijing Oligobi0 Biotechnology Co., Ltd. (Beijing, China) to synthesize shRNA. The DNA sequences corresponding to the shRNA specifically targeting the BTF3L4 gene are shown below:

[0065] BTF3L4-sh1: 5′-CCTGATGTTACAGTTTGGTAGATTT-3′,

[0066] BTF3L4-sh2: 5′-CAAACTGGAATAGCTAGCATGTGCT-3′.

[0067] 2. Culture of Glioma Cell Lines

[0068] Glioma cell lines, including: T98G cells, U87mg cells, U251 cells, SHG44 cells, are cultured in DMEM complete medium containing 10% fetal bovine serum. The temperature is maintained at 37°C and the CO2 humidity is 5% in the incubator. Regular passage culture is carried out, and the medium is changed every 2 - 3 days. Cells in the logarithmic growth phase are selected for experiments.

[0069] 3. Expression of BTF3L4 in Glioma Tissues and Cell Lines

[0070] 1) Extraction of Glioma Tissue Proteins

[0071] Add 1 mL of pre-cooled protein lysis buffer to every 250 mg of tissue; place it in a tissue ultrasonic lyser for sufficient lysis, with an ice bath for 1 minute between each lysis until the tissue is completely lysed; place the tissue homogenate in a pre-cooled centrifuge and centrifuge at 12000g for 15 minutes, then immediately transfer the supernatant into a new centrifuge tube for storage and later use.

[0072] 2) Extraction of Total Cellular Proteins

[0073] Take out the corresponding cells from the 37°C incubator; discard the medium, wash the cells 2 times with pre-cooled PBS, discard the PBS and suck up the remaining PBS solution to avoid diluting cellular proteins; according to the size of the cell culture flask and the growth density of the cells, add RIPA cell lysis buffer, then use a cell scraper to collect the cells and transfer them to a clean EP tube; fully lyse the scraped cellular proteins on ice for 20 - 30 minutes; centrifuge at 12000 rpm at 4°C for 15 min; take the supernatant, measure the concentration of cellular proteins by the BCA method, then add loading buffer, pipette and mix well, boil at 95°C for 5 minutes, aliquot, and store in a -80°C refrigerator for later use.

[0074] 3) Western blot

[0075] Prepare polyacrylamide gels (5% stacking gel, 12.5% separating gel); clean the glass plates, let them dry tilted, assemble the glass plates, add the separating gel, fill it up to 2 cm from the upper end of the glass plates, immediately add isopropanol for liquid sealing, let it stand for 30 min. After the separating gel solidifies, gently pour out the upper layer of isopropanol, then add the stacking gel to the top of the glass plates, immediately insert the comb, and let it stand for 30 min until the stacking gel solidifies; put the prepared gel into the electrophoresis tank, fill it up with electrophoresis buffer. After loading Protein Maker and the extracted protein samples, add the remaining electrophoresis solution, connect the power supply, adjust the voltage to 80 V. After the Protein marker separates, then adjust the voltage to 100 V. After completion, take out the gel and cut the target band; cut a PVDF membrane of appropriate size, polarize it in methanol for about 30 s first, and then put it into the transfer buffer; at the same time, soak the sponge and filter paper in the transfer buffer for 20 min, install the transfer device, and the discharge order is: cathode carbon plate + sponge + filter paper + gel + PVDF membrane + filter paper + sponge + anode carbon plate; put the transfer device into the transfer tank, add ice bags, and fill it up with transfer buffer; connect the power supply, adjust the wet transfer at a constant current of 300 mA for 0.5 h, and the transfer process needs to be carried out in an ice box throughout; after the transfer is completed, put the PVDF membrane into the blocking solution (5 g of skim milk powder dissolved in 100 mL of TBST), block it at 80 r / min at room temperature on a shaker for 2 h; after blocking, prepare the primary antibody dilution solution with the blocking solution according to the primary antibody dilution ratio, evenly drip the diluted primary antibody on the PVDF membrane, and incubate overnight at 4°C; the next day, wash the membrane 3 times with TBST, 15 min each time; after washing the membrane, prepare the secondary antibody dilution solution with TBST according to the secondary antibody dilution ratio, evenly drip the diluted secondary antibody on the PVDF membrane, and incubate at room temperature for 1.5 h; after incubation, wash the membrane 3 times with TBST, 15 min each time; after washing the membrane, dry the PVDF membrane with filter paper, lay it flat at the corresponding position of the imaging instrument. ECL luminescent solution is mixed with solution A and solution B in equal proportions before use, and evenly dripped on the PVDF membrane, and take a photo and save it with a gel imaging system.

[0076] Extract the proteins of 6 pairs of surgically resected glioma tissues and adjacent tissues and 4 types of glioma cells respectively according to the above method. Detect the expression of BTF3L4 in 4 types of glioma cells by Western blot, and screen out the high-expression and low-expression cells. The results are as Figure 4 shown. The expression of BTF3L4 protein in glioma tissues is higher than that in its corresponding adjacent tissues, and the expression of BTF3L4 protein is relatively high in U251 glioma cells and relatively low in U87mg glioma cells.

[0077] 4. Interference of BTF3L4 gene expression in glioma cells by the RNAi system

[0078] Using the constructed RNAi interference system, specifically transfect U251 glioma cells with the BTF3L4-sh1 sequence: 5′-CCTGATGTTACAGTTTGGTAGATTT-3′; Inoculate the target cells to be transfected in a 12-well plate. When the cells are completely spread out, the confluence reaches 60 - 70%, and the cell state is good, transfection of the cells can be started; After 6 - 8 hours of infection, change the medium and continue culturing. After 48 hours, collect the cells, lyse them and extract proteins, and use Western blot to detect the transfected U251 cells.

[0079] The results are as Figure 5 shown. Compared with the untreated group, the relative expression level of BTF3L4 protein after RNAi interference was significantly reduced, indicating that the expression of BTF3L4 protein was effectively inhibited.

[0080] 5. Cell migration (Transwell chamber method)

[0081] Digest and collect the cells of each group 48 h after transfection, centrifuge and set aside; Resuspend the cells with basal medium and adjust the cell density to 5×10 4 / mL; Add 800 μL of complete medium to a 24-well plate, place the chamber, soak it thoroughly, take 100 μL of the cell suspension and add it to the upper chamber; After culturing routinely for 24 h, take it out, wash it 2 times with 1×PBS, fix it with 4% paraformaldehyde for 20 min, and wash it 2 times with 1×PBS; Add 500 μL of crystal violet staining solution to the 24-well plate, place the chamber, take it out after 10 min, wash it 2 times with 1×PBS, invert the chamber, and gently wipe off the cells that did not pass through the upper chamber with a cotton swab.

[0082] The results are as Figure 6 shown. The migration ability of U251 cells decreased after interfering with BTF3L4.

[0083] 6. Cell invasion (Transwell chamber method)

[0084] First, prepare Matrigel matrix glue (BD Biosciences, San Jose, CA), add 100 μL to each upper chamber of the Transwell chamber, avoiding air bubbles; Digest and collect the cells of each group 48 h after transfection, centrifuge and set aside; Resuspend the cells with basal medium and adjust the cell density to 5×10 4 / mL; Add 800 μL of complete medium to a 24-well plate, place the insert, soak it thoroughly, and transfer 100 μL of cell suspension to the upper chamber; After culturing routinely for 24 - 48 h, take it out, wash twice with 1×PBS, fix with 4% paraformaldehyde for 20 min, and wash twice with 1×PBS; Add 500 μL of crystal violet staining solution to the 24-well plate, place the insert, take it out after 10 min, wash twice with 1×PBS, invert the insert, and gently wipe off the cells that did not pass through the upper chamber with a cotton swab.

[0085] The results are as Figure 7 shown. After interfering with BTF3I4, the invasion ability of U251 cells decreased.

[0086] 7. Cell cycle detection

[0087] After the cells were confluent, adjust the cell concentration to 10 5 cells / mL, inoculate into a 6-well culture plate, with 3 mL of culture medium in each well, and culture in a 37 °C, 5% CO2 incubator for 24 hours; Continue to culture for 48 hours after transfection with siRNA-1; Digest the cells in the plate with trypsin to prepare a single-cell suspension, and stain with a cell cycle detection kit (C1052, Beyotime) after fixation. Subsequently, use a BD-FACSVerse flow cytometer to detect the percentage of cells in different division stages, and finally perform quantitative analysis with CELL Quest software.

[0088] The results are as Figure 8 shown. After interfering with BTF3L4, the quantitative results showed that the number of U251 glioma cells in the S phase increased significantly, and the growth of glioma cells in the G0 / G1 phase (2N) and G2 / M phase (4N) was significantly blocked, and the cell differentiation ability was impaired.

Claims

1. BTF3L4 Use of an RNAi interference system of a gene in the preparation of a drug for treating glioma, characterized in that, The described BTF3L4 The sequences of the RNAi interference system of the gene are as follows: BTF3L4 -sh1: 5'-CCTGATGTTACAGTTTGGTAGATTT-3', BTF3L4 -sh2: 5'-CAAACTGGAATAGCTAGCATGTGCT-3'.

2. The use according to claim 1, characterized in that, The BTF3L4 RNAi interference system of the gene is to knockdown the BTF3L4 gene in glioma tissue cells.

3. The use according to claim 1, characterized in that, The BTF3L4 RNAi interference system of the gene is to inhibit BTF3L4 protein expression in glioma tissue cells.

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