Use of gemin 4 gene and gemin 4 protein in tumor

Abstract

The present invention relates to the technical field of biomedicine, and provides a use of a Gemin 4 gene and a Gemin 4 protein in a tumor. The present invention specifically relates to a use of a Gemin 4 gene and a Gemin 4 protein in the preparation of a reagent or a drug for detecting, preventing or treating a tumor, and a use of the Gemin 4 protein in the preparation of a reagent for detecting a tumor. The tumor comprises one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

Description

Application of Gemin4 gene and Gemin4 protein in tumors Technical Field

[0001] The present application belongs to the field of biomedicine technology, and specifically relates to the application of Gemin4 gene and Gemin4 protein in tumors. Background Art

[0002] Malignant tumors are the leading cause of death in humans. Unlimited growth, invasion, and metastasis are hallmarks of malignancy and the leading cause of treatment failure and death. Currently, the effectiveness of drugs is severely limited by the high metastatic potential of tumors and the heterogeneity of tumor cells. Continuous research into the causes of tumors, the development of treatment and prevention strategies, and the development of new and effective drugs are key to clinical cancer treatment.

[0003] Tumor markers are substances produced by tumor cells themselves or abnormally produced and / or elevated in response to tumor cells during the development and proliferation of malignant tumors, reflecting the presence and / or growth of the tumor. These substances include proteins, hormones, enzymes (isoenzymes), polyamines, and oncogene products. Found in a patient's blood, body fluids, cells, or tissues, they can be measured using biochemical, immunological, and molecular biological methods. They are valuable in assisting the diagnosis, differential diagnosis, therapeutic efficacy observation, recurrence monitoring, and prognostic assessment of tumors. Currently, the number of markers that can be used for precise tumor diagnosis is very limited. Discovering more effective tumor detection indicators and improving the diagnostic value of tumor markers are urgent challenges that need to be addressed.

[0004] Gemin4 (Gem Nuclear Organelle Associated Protein 4) is a key member of the GEMIN protein family. As a crucial component of the survival motor neuron (SMN) complex, it maintains the structural integrity of small nuclear ribonucleoproteins (snRNPs) by binding to small nuclear RNAs (snRNAs) with motor neuron proteins (SMNs) and GEMIN2, 3, 5, 6, 7, and 8 within the complex, thereby promoting spliceosome assembly and downstream pre-mRNA activation. Previous studies have shown that mutations in genes encoding components of the SMN complex are closely associated with the motor neuron disease spinal muscular atrophy (SMA). Furthermore, abnormal expression of Gemin5 and Gemin6 is associated with the malignant phenotype of various cancers. Recent studies have linked GEMIN4 gene polymorphisms to the development of colon, bladder, liver, kidney, and ovarian cancers. However, the role of GEMIN4 gene expression levels and Gemin4 protein expression in different tumors remains unclear.

[0005] Summary of the Invention

[0006] 1. Purpose of the Invention

[0007] One of the purposes of this application is to provide the use of the Gemin4 gene in the preparation of tumor detection reagents, and drugs for preventing or treating tumors.

[0008] The second purpose of this application is to provide the use of Gemin4 protein in the preparation of tumor detection reagents.

[0009] The third purpose of this application is to provide a reagent for detecting the expression level of the Gemin4 gene and a reagent for detecting the protein level of Gemin4 for use in the preparation of tumor detection products.

[0010] The fourth purpose of the present application is to provide a reagent for knocking down or silencing Gemin4 gene expression for use in the preparation of drugs for preventing or treating tumors.

[0011] 2. Technical solution

[0012] In order to achieve the above-mentioned invention objectives, the technical solutions adopted in this application are as follows:

[0013] The present application provides the use of the Gemin4 gene in preparing tumor detection reagents, and drugs for preventing or treating tumors. The nucleic acid sequence of the Gemin4 gene is shown in SEQ ID NO.1.

[0014] Furthermore, the above-mentioned tumors include one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

[0015] Furthermore, the Gemin4 gene is highly expressed in human glioma, liver cancer, or colorectal cancer.

[0016] Furthermore, the Gemin4 gene is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, or lung cancer.

[0017] Furthermore, the above tumor detection reagent includes a specific primer pair with nucleotide sequences as shown in SEQ ID NO.3 and SEQ ID NO.4.

[0018] Furthermore, the above-mentioned prevention or treatment of tumors is achieved by knocking down or silencing the expression of Gemin4 gene in tumor cells.

[0019] Furthermore, the knockdown or silencing of Gemin4 gene expression is achieved by designing specific small interfering RNA according to the nucleotide sequence of the Gemin4 gene and transfecting the specific small interfering RNA.

[0020] Furthermore, the nucleotide sequence of the above-mentioned specific small interfering RNA is shown in SEQ ID NO: 5.

[0021] Furthermore, the above-mentioned prevention or treatment of tumors includes inhibiting the growth and / or metastasis of colorectal cancer, liver cancer and glioma cells.

[0022] Furthermore, the tumor cells of the above tumor include one or more of the following:

[0023] Glioma cancer cells include: U87 and / or T98G;

[0024] Colorectal cancer cells include: HT29;

[0025] Liver cancer cells include: HepG2.

[0026] The present application also provides the use of Gemin4 protein in preparing a tumor detection reagent. The amino acid sequence of the Gemin4 protein is shown in SEQ ID NO.2.

[0027] Furthermore, the above-mentioned tumors include one or more of human head and neck cancer, lung cancer, gastric cancer, bile duct cancer, breast cancer, kidney cancer, esophageal cancer, colorectal cancer, liver cancer and glioma.

[0028] Furthermore, Gemin4 protein is highly expressed in human glioma, liver cancer or colorectal cancer.

[0029] Furthermore, Gemin4 protein is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer or lung cancer.

[0030] The present application also provides the use of a reagent for detecting the expression level of the Gemin4 gene in the preparation of a tumor detection product.

[0031] Furthermore, the above-mentioned tumors include one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

[0032] Furthermore, the reagent for detecting the expression level of the Gemin4 gene is a specific primer pair designed based on the nucleotide sequence of the Gemin4 gene as shown in SEQ ID NO.1.

[0033] Furthermore, the nucleotide sequences of the above-mentioned specific primer pair are shown in SEQ ID NO.3 and SEQ ID NO.4.

[0034] Furthermore, the Gemin4 gene is highly expressed in human glioma, liver cancer, or colorectal cancer.

[0035] Furthermore, the Gemin4 gene is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, or lung cancer.

[0036] The present application also provides the use of a reagent for detecting Gemin4 protein levels in the preparation of a tumor detection product.

[0037] Furthermore, the above-mentioned tumors include one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

[0038] Furthermore, the reagent for detecting the level of Gemin4 protein includes a specific antibody for Gemin4 protein, and can be detected by ELISA or other means.

[0039] Furthermore, Gemin4 protein is highly expressed in human glioma, liver cancer or colorectal cancer.

[0040] Furthermore, Gemin4 protein is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer or lung cancer.

[0041] The present application also provides a tumor detection kit, comprising the above-mentioned reagent for detecting the Gemin4 gene expression level or the above-mentioned reagent for detecting the Gemin4 protein level.

[0042] Furthermore, the above-mentioned tumors include one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

[0043] Furthermore, the reagent for detecting the expression level of the Gemin4 gene is a specific primer pair designed based on the nucleotide sequence of the Gemin4 gene as shown in SEQ ID NO.1.

[0044] Furthermore, the nucleotide sequences of the above-mentioned specific primer pair are shown in SEQ ID NO.3 and SEQ ID NO.4.

[0045] Furthermore, the Gemin4 gene is highly expressed in human glioma, liver cancer, or colorectal cancer.

[0046] Furthermore, the Gemin4 gene is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, or lung cancer.

[0047] Furthermore, the reagent for detecting the level of Gemin4 protein includes a specific antibody for Gemin4 protein, and can be detected by ELISA or other means.

[0048] Furthermore, Gemin4 protein is highly expressed in human glioma, liver cancer or colorectal cancer.

[0049] Furthermore, Gemin4 protein is lowly expressed in human esophageal cancer, renal cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer or lung cancer.

[0050] The present application also provides the use of an agent for knocking down or silencing Gemin4 gene expression in the preparation of a drug for preventing or treating tumors.

[0051] Furthermore, the above-mentioned tumors include one or more of glioma, colorectal cancer, and liver cancer.

[0052] Furthermore, the above-mentioned prevention or treatment of tumors is achieved by knocking down or silencing the expression of Gemin4 gene in tumor cells.

[0053] Furthermore, the knockdown or silencing of Gemin4 gene expression is achieved by designing specific small interfering RNA based on the nucleotide sequence of the Gemin4 gene.

[0054] Furthermore, the reagent for knocking down or silencing Gemin4 gene expression includes specific siRNA targeting the Gemin4 gene.

[0055] Furthermore, the reagent for knocking down or silencing Gemin4 gene expression includes siRNA having a nucleotide sequence as shown in SEQ ID NO.5.

[0056] Furthermore, the above-mentioned prevention or treatment of tumors includes inhibiting tumor cell growth and / or metastasis.

[0057] Furthermore, the tumor cells of the above tumor include one or more of the following:

[0058] Glioma cancer cells include: U87 and / or T98G;

[0059] Colorectal cancer cells include: HT29;

[0060] Liver cancer cells include: HepG2.

[0061] The present application also provides a pharmaceutical composition for preventing or treating tumors, which contains at least a reagent for knocking down or silencing Gemin4 gene expression and a pharmaceutically acceptable carrier.

[0062] Furthermore, the above-mentioned tumors include one or more of glioma, colorectal cancer, and liver cancer.

[0063] Furthermore, the reagent for knocking down or silencing Gemin4 gene expression includes specific siRNA targeting the Gemin4 gene.

[0064] Furthermore, the reagent for knocking down or silencing Gemin4 gene expression includes siRNA having a nucleotide sequence as shown in SEQ ID NO.5.

[0065] Furthermore, the above-mentioned prevention or treatment of tumors includes inhibiting tumor cell growth and / or metastasis.

[0066] Furthermore, the tumor cells of the above tumor include one or more of the following:

[0067] Glioma cancer cells include: U87 and / or T98G;

[0068] Colorectal cancer cells include: HT29;

[0069] Liver cancer cells include: HepG2.

[0070] 3. Beneficial effects

[0071] Compared with the prior art, the present application has the following advantages:

[0072] (1) This application uses a fluorescent quantitative PCR method to detect the expression level of the Gemin4 gene in tumor cells and normal cells. It is found that the expression level of the Gemin4 gene is significantly reduced in human esophageal cancer cells TE13 and Eca109, renal cancer cells A498, breast cancer cells MDA-MB-231, gastric cancer cells MGC803, head and neck cancer cells CAL27, bile duct cancer cells RBE and HuCC-T1, and lung cancer cells A549; and the expression level is significantly increased in human glioma cells T98G and U87, liver cancer cells HepG2, and colorectal cancer cells HT29. The above findings indicate that the Gemin4 gene can be used as a potential tumor marker for auxiliary diagnosis of early tumors.

[0073] (2) Based on the fluorescent quantitative PCR method to verify the expression level of Gemin4 gene in tumor cells, the present application used a specific antibody of Gemin4 protein for immunoblotting detection, and found that the results were consistent with the qPCR results. The expression level of Gemin4 protein in human esophageal cancer cells TE13 and Eca109, renal cancer cells A498, breast cancer cells MDA-MB-231, gastric cancer cells MGC803, head and neck cancer cells CAL27, bile duct cancer cells RBE and HuCC-T1, and lung cancer cells A549 was significantly reduced; the expression level in human glioma cells T98G and U87, liver cancer cells HepG2, and colorectal cancer cells HT29 was significantly increased. The above findings indicate that Gemin4 protein can be used as a potential tumor marker for auxiliary diagnosis of early tumors.

[0074] (3) The present application designs specific siRNA targeting the nucleotide sequence encoding Gemin4 protein for human glioma cells T98G and U87, liver cancer cells HepG2, and colorectal cancer cells HT29, which have significantly elevated Gemin4 gene expression levels. After transfection of the above cancer cells, it was found that the growth and / or metastasis of glioma, liver cancer, and colorectal cancer cells can be significantly inhibited, indicating that targeted inhibition of the Gemin4 gene (knockdown or silencing) can be used to treat glioma, liver cancer, and colorectal cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] Figure 1 shows the situation in which qPCR detected that the Gemin4 gene was significantly underexpressed in different tumor cells.

[0076] Figure 2 shows the results of qPCR detection of significantly high expression of the Gemin4 gene in different tumor cells.

[0077] Figure 3 shows the significantly low expression of Gemin4 protein in different tumor cells detected by Western Blot.

[0078] FIG4 shows the results of Western Blot detection of Gemin4 protein that is significantly overexpressed in different tumor cells.

[0079] FIG5 shows the expression of Gemin4 gene at RNA and protein levels after transfection of si-Gemin4.

[0080] FIG6 shows the inhibitory effect of knocking down the Gemin4 gene on tumor cell proliferation.

[0081] FIG7 shows the inhibitory effect of knocking down the Gemin4 gene on tumor cell migration. DETAILED DESCRIPTION

[0082] The present application is further described below with reference to specific embodiments.

[0083] It should be noted that the terms such as "upper", "lower", "left", "right", and "middle" cited in this specification are only for the convenience of description and are not used to limit the scope of implementation. Changes or adjustments to their relative relationships should be regarded as the scope of implementation of this application without substantially changing the technical content.

[0084] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the term "and / or" used herein includes any and all combinations of one or more of the associated listed items.

[0085] If the specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or those recommended by the manufacturer. All reagents or instruments used, if the manufacturer is not specified, are commercially available conventional products.

[0086] As used herein, the term "about" is used to provide flexibility and imprecision associated with a given term, measurement, or value. One skilled in the art can readily determine the degree of flexibility for a particular variable.

[0087] As used herein, the term "at least one of" is intended to be synonymous with "one or more of." For example, "at least one of A, B, and C" explicitly includes only A, only B, only C, and combinations of each thereof.

[0088] Concentration, amount and other numerical data can be presented in range format in this article.Should be understood that such range format is only used for convenience and brevity, and should be flexibly interpreted as not only including the numerical value clearly described as range limit, but also including all independent numerical values ​​or subranges encompassed within the scope, just as each numerical value and subrange are clearly described.For example, the numerical range of about 1 to about 4.5 should be interpreted as not only including the limit value of 1 to about 4.5 clearly described, but also including independent numerals (such as 2,3,4) and subranges (such as 1 to 3,2 to 4 etc.).The same principle is applicable to the scope of only narrating a numerical value, such as "less than about 4.5", which should be interpreted as including all above-mentioned values ​​and scopes.In addition, no matter how the breadth of described scope or feature is, this explanation should be applicable.

[0089] Example 1

[0090] In this example, the fluorescence quantitative PCR method was used to detect the expression level of the Gemin4 gene in tumor cells and normal cells.

[0091] Tumor cells include:

[0092] Human microglial cells HMC3, human glioma cells T98G and U87;

[0093] Human hepatocytes L-02 and HepG2;

[0094] Colorectal cancer cells HT29;

[0095] Human normal esophageal epithelial cells HEEC, esophageal cancer cells TE13 and Eca109;

[0096] Human renal epithelial cells HEK293T and renal cancer cells A498;

[0097] Human normal mammary epithelial cells MCF10A and breast cancer cells MDA-MB-231;

[0098] Gastric cancer cells MGC803;

[0099] Head and neck cancer cell CAL27;

[0100] Cholangiocarcinoma cells RBE and HuCCT1;

[0101] Lung cancer A549 cells.

[0102] Detection methods include:

[0103] (1) Extraction of total RNA from tumor cells and normal epithelial cells

[0104] The above 18 cell types were cultured in an incubator at 37°C and 5% CO2. When the density reached 90%, the cells were digested and collected by trypsinization. The cells were resuspended in culture medium and counted under a microscope. The cell concentration was adjusted to 5×10 5 The cell suspension with adjusted concentration was then inoculated into 6-well plates with 2 mL per well, and cultured in a 37°C, 5% CO2 incubator for 24 h.

[0105] Total RNA of the above cells was extracted according to the Trizol instructions of Tiangen Bio. The purity and concentration of the extracted RNA were quantified using NanoDrop ND-1000 nucleic acid quantifier, and the integrity of the extracted RNA was ensured by agarose quality inspection.

[0106] (2) RNA reverse transcription to synthesize first-strand cDNA

[0107] The extracted total RNA was reverse transcribed into cDNA using the ABM PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time). This kit contains gDNA Eraser DNase, which effectively removes contaminated genomic DNA.

[0108] (3) Real-time quantitative PCR

[0109] Specific primers were designed based on the nucleic acid sequences of Gemin4 and GAPDH genes, and the ABM kit was used to Premix Ex Taq TM II (TliRNaseH Plus) for qPCR reaction. The upstream and downstream primer sequences of Gemin4 are shown as SEQ ID NO.3 and SEQ ID NO.4, respectively. The upstream and downstream primer sequences of GAPDH are shown as SEQ ID NO.6 and SEQ ID NO.7, respectively. The reaction system is shown in Table 1:

[0110] Table 1 PCR reaction system

[0111] After mixing the above components evenly, real-time quantitative PCR was performed according to the following program: pre-denaturation at 95°C for 30 s, 40 cycles; 95°C for 5 s, 60°C for 30 s.

[0112] The specificity of the reaction was determined based on the melting curve, and the formula 2 -ΔΔCt The expression level of the Gemin4 gene was calculated. The results are shown in Figures 1 and 2. Figure 1 shows that compared with normal human cells, the expression level of the Gemin4 gene was significantly decreased in human esophageal cancer cells TE13 and Eca109, renal cancer cells A498, breast cancer cells MDA-MB-231, gastric cancer cells MGC803, head and neck cancer cells CAL27, bile duct cancer cells RBE and HuCC-T1, and lung cancer cells A549; Figure 2 shows that compared with normal human cells, the expression level of the Gemin4 gene was significantly increased in human glioma cells T98G and U87, liver cancer cells HepG2, and colorectal cancer cells HT29.

[0113] Example 2

[0114] In this example, the Western Blot method was used to detect the expression level of Gemin4 protein in tumor cells and normal cells.

[0115] Tumor cells include:

[0116] Human microglial cells HMC3, human glioma cells T98G and U87;

[0117] Human hepatocytes L-02 and HepG2;

[0118] Colorectal cancer cells HT29;

[0119] Human normal esophageal epithelial cells HEEC, esophageal cancer cells TE13 and Eca109;

[0120] Human renal epithelial cells HEK293T and renal cancer cells A498;

[0121] Human normal mammary epithelial cells MCF10A and breast cancer cells MDA-MB-231;

[0122] Gastric cancer cells MGC803;

[0123] Head and neck cancer cell CAL27;

[0124] Cholangiocarcinoma cells RBE and HuCCT1;

[0125] Lung cancer A549 cells.

[0126] Detection methods include:

[0127] Protein extraction from tumor cells and normal epithelial cells:

[0128] The above 18 cell types were cultured in an incubator at 37°C and 5% CO2. When the density reached 90%, the cells were digested and collected by trypsinization. The cells were resuspended in culture medium and counted under a microscope. The cell concentration was adjusted to 5×10 5 The cell suspension with adjusted concentration was then inoculated into 6-well plates with 2 mL per well, and cultured in a 37°C, 5% CO2 incubator for 24 h.

[0129] After centrifugation, discard the supernatant, rinse twice with PBS, and discard the supernatant. Add RIPA lysis buffer and lyse on ice for 20 minutes. Centrifuge at 12,000 g for 10 minutes and collect the supernatant. Add 1× SDS loading buffer, mix thoroughly by pipetting, and then denature by boiling for 5 minutes. Separate total proteins on a 10% SDS-PAGE gel and transfer to a PVDF membrane. Block with 5% BSA at room temperature for 2 hours, incubate with GAPDH and Gemin4 antibodies overnight at 4°C, respectively, and wash three times with TBST. Incubate with secondary antibodies at room temperature for 1 hour and wash three times with TBST. Develop with ECL ultrasensitive chemiluminescence solution and detect the presence of target bands using a Tanon imaging system.

[0130] The results are shown in Figures 3 and 4. Figure 3 shows that compared with normal human cells, the expression of Gemin4 protein in human esophageal cancer cells TE13 and Eca109, renal cancer cells A498, breast cancer cells MDA-MB-231, gastric cancer cells MGC803, head and neck cancer cells CAL27, bile duct cancer cells RBE and HuCC-T1, and lung cancer cells A549 is significantly reduced; as shown in Figure 4, Figure 4 shows that compared with normal human cells, the expression of Gemin4 protein in human glioma cells T98G and U87, liver cancer cells HepG2, and colorectal cancer cells HT29 is significantly increased. The above results are consistent with the qPCR results of Example 1, indicating that Gemin4 can be used as a potential marker for tumor detection.

[0131] Example 3

[0132] This example provides the effect of silencing Gemin4 gene expression on the proliferation ability of tumor cells.

[0133] Specific siRNA was designed for the nucleotide sequence of the Gemin4 gene. The specific sequence is shown in SEQ ID NO.5 (GCUCCUGUGUGAGAUUGUATT). Glioma cells U87, colorectal cancer cells HT29, and liver cancer cells HepG2 were cultured in a 37°C, 5% CO2 incubator. The cells were digested 24 hours in advance and plated in 12-well plates at a density of 8×104 / mL, and cell transfection was performed when the cell confluence reached 60%. Add 50μL DMEM culture medium to a 1.5mL sterile EP tube, add 8μL transfection reagent, pipette to mix, and let it stand at room temperature for 5min. Add 50μL DMEM culture medium to the other two EP tubes, add 6μL siRNA (si-Gemin4 and si-NC, the nucleotide sequence of si-NC is shown in SEQ ID NO.8: ACGUGACACGUUCGGAGAATT), pipette to mix, and let it stand for 5min. Add the transfection reagent mixture obtained in the first step dropwise to the second EP tube, pipette to mix, and incubate at room temperature for 20min. While incubating, aspirate the original culture medium in the 6-well plate and replace it with fresh complete culture medium. After the incubation is completed, add the transfection mixture dropwise to the 6-well plate, shake to mix, and place it in the incubator for culture. Change the medium after 6h, and digest and carry out functional experiments after 48h.

[0134] The two groups of transfected cells were cultured in a 37°C, 5% CO2 incubator until the density reached above 90%. The cells were then digested and collected using trypsin. The cells were resuspended in culture medium and counted under a microscope. The cell concentration was adjusted to 3.0×10 4 Cell suspensions were plated into 96-well plates at a concentration of 100 μL per well and incubated in a 37°C, 5% CO2 incubator for 48 hours. In a dark-protected environment, 10 μL of CCK8 reagent was added to each well and incubated in a 37°C, dark-protected water bath for 4 hours. A microplate reader was preheated 30 minutes in advance. After incubation, the absorbance of each well was measured at 450 nm. Data were analyzed. The experiment was repeated three times. Results are expressed as mean ± SD and statistically analyzed using a T-test. *P < 0.05 indicates significant differences, and **P < 0.01 indicates extremely significant differences.

[0135] The results are shown in Figure 5. si-Gemin4 successfully silenced the expression of the Gemin4 gene at the RNA and protein levels. Silencing the Gemin4 gene significantly inhibited the proliferation of glioma cells U87, colorectal cancer cells HT29, and liver cancer cells HepG2 (Figure 6). This indicates that targeted inhibition of the Gemin4 gene can exert an anti-tumor effect by inhibiting the proliferation of tumor cells.

[0136] Example 4

[0137] This example provides the effect of silencing Gemin4 gene expression on the migration ability of tumor cells.

[0138] The three tumor cells (glioma cells U87, colorectal cancer cells HT29, and liver cancer cells HepG2) transfected with si-Gemin4 and si-NC in Example 3 were digested, counted, and inoculated into transwell chambers, 100 μL per well. 0.6 mL of complete culture medium containing 10% FBS was then added to the lower chamber of the transwell to stimulate cell migration, and the cells were cultured at 5% CO2 and 37°C for 48 h. The culture medium in the wells was discarded, the cells were fixed with methanol at room temperature for 30 min, stained with 0.1% crystal violet at room temperature for 10 min, rinsed with clean water, and the unmigrated cells on the upper layer were wiped off with a cotton swab. The cells were observed under a microscope and four fields of view were selected for photographing and counting. The experiment was repeated 3 times independently. The results obtained in the experiment are expressed as mean ± SD, and statistical T test was performed. *P < 0.05 indicates a significant difference, and **P < 0.01 indicates an extremely significant difference.

[0139] The results are shown in Figure 7. Silencing Gemin4 gene expression can significantly inhibit the migration of glioma cells U87, colorectal cancer cells HT29, and liver cancer cells HepG2, indicating that targeted inhibition of the Gemin4 gene can exert an anti-tumor effect by inhibiting the metastasis of tumor cells.

[0140] The experiments designed in the present invention are scientifically sound, feasible, and effective. Based on the above findings, the Gemin4 gene expression level or Gemin4 protein level can be used as a new biomarker to assist in the diagnosis of malignant tumors including glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer. The Gemin4 gene and / or Gemin4 protein can be directly used as a therapeutic drug for malignant tumors or as a potential target for anti-tumor drugs, greatly expanding the use of the gene and / or protein and providing new ideas and prospects for future drug development.

Claims

1. The use of Gemin4 gene in the preparation of tumor detection reagents, tumor prevention or treatment drugs, characterized in that: The nucleic acid sequence of the gene is shown in SEQ ID NO.

1.

2. The use of the Gemin4 gene according to claim 1 in the preparation of tumor detection reagents, tumor prevention or treatment drugs, characterized in that: The tumor includes one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

3. The use of the Gemin4 gene according to claim 2 in preparing a tumor detection reagent, characterized in that: The tumor detection reagent includes a specific primer pair with nucleotide sequences as shown in SEQ ID NO.3 and SEQ ID NO.

4.

4. The use of the Gemin4 gene according to claim 1 or 2 in preparing a drug for preventing or treating tumors, characterized in that: The prevention or treatment of tumors comprises knocking down or silencing the expression of Gemin4 gene in tumor cells.

5. The use of the Gemin4 gene according to claim 4 in preparing a drug for preventing or treating tumors, characterized in that: The knocking down or silencing of Gemin4 gene expression comprises transfecting a specific small interfering RNA designed according to the nucleotide sequence of the Gemin4 gene, wherein the nucleotide sequence of the specific small interfering RNA is shown in SEQ ID NO:

5.

6. Use of the Gemin4 gene according to claim 4 or 5 in preparing a drug for preventing or treating tumors, characterized in that: The prevention or treatment of tumors includes inhibiting the growth and / or metastasis of colorectal cancer, liver cancer and glioma cells.

7. Application of Gemin4 protein in the preparation of tumor detection reagents, characterized in that: The amino acid sequence of the Gemin4 protein is shown in SEQ ID NO.

2.

8. The use of the Gemin4 protein in the preparation of a tumor detection reagent according to claim 7, characterized in that: The tumor includes one or more of human head and neck cancer, lung cancer, gastric cancer, bile duct cancer, breast cancer, kidney cancer, esophageal cancer, colorectal cancer, liver cancer and glioma.

9. A tumor detection kit, characterized in that: It includes reagents for detecting the expression level of Gemin4 gene or reagents for detecting the level of Gemin4 protein, and the tumor includes one or more of human glioma, liver cancer, colorectal cancer, esophageal cancer, kidney cancer, breast cancer, gastric cancer, head and neck cancer, bile duct cancer, and lung cancer.

10. A pharmaceutical composition for preventing or treating tumors, characterized in that: The pharmaceutical composition for preventing or treating tumors contains at least a reagent for knocking down or silencing Gemin4 gene expression and a pharmaceutically acceptable carrier, and the tumors include one or more of glioma, colorectal cancer, and liver cancer.

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