Application of PELI2 gene as renal clear cell carcinoma molecular marker

Abstract

The invention relates to application of a PELI2 gene as a molecular marker of renal clear cell carcinoma. The nucleotide sequence of the PELI2 gene is as shown in SEQ ID NO. 1. It is found for the first time that the PELI2 gene is remarkably low in expression in renal clear cell carcinoma cells, and the expression level of the PELI2 gene is closely related to prognosis of patients. The expression of the PELI2 gene in the renal clear cell carcinoma cells is remarkably reduced, and the survival rate of a renal cancer patient with high expression of the PELI2 gene is remarkably higher than that of a renal cancer patient with low expression of the PELI2 gene. Meanwhile, experiments prove that the proliferation capacity of renal clear cell carcinoma cells can be inhibited by overexpression of the PELI2 gene. On the basis, a primer pair capable of specifically recognizing the PELI2 gene and an overexpression lentivirus of the PELI2 gene are designed, and the application of the PELI2 gene as a marker in diagnosis and prognosis evaluation of the renal clear cell carcinoma and targeted therapy of the renal clear cell carcinoma is provided, which is of great significance in subsequent drug research of the renal clear cell carcinoma.

Description

TECHNICAL FIELD

[0001] The application relates to application of a PELI2 gene as a molecular marker of renal clear cell carcinoma and belongs to the technical field of biomedicine. BACKGROUND

[0002] Renal cell carcinoma (RCC) has been listed as the seventh cancer type in developed countries, affecting more than 400,000 people worldwide. RCC has several subtypes, and the most common pathological and lethal type is renal clear cell carcinoma (KIRC), which accounts for about 70-80% of RCC, and surgery and targeted immunotherapy are indispensable treatment options for renal cell carcinoma. However, tumor recurrence and drug resistance are major problems in the treatment of RCC. Therefore, screening of effective biomarkers has important clinical significance for the diagnosis, treatment and prognosis evaluation of KIRC.

[0003] Pellino-2 (PELI2) is a newly discovered E3 ubiquitin ligase that regulates protein degradation, protein-protein interaction, protein translocation and signal transduction through ubiquitination of target proteins. PELI2 has a C-terminal ring-like domain and a phospho-threonine binding forkhead associated (FHA) domain, which are responsible for ubiquitin ligase activity and substrate binding, respectively. More and more evidence shows that PELI2 acts as a key mediator of innate immunity and inflammation through multiple signaling of IL-1 receptor, Toll-like receptor and NOD-like receptor. In addition, it is reported that PELI2 plays an important role in lung injury and cancer. However, so far there is no relevant report on the role of PELI2 gene in KIRC.

[0004] At present, the research on the pathogenesis, disease diagnosis and treatment of PELI2 gene in renal clear cell carcinoma is still very insufficient, therefore, the role of PELI2 in the pathogenesis, disease diagnosis and treatment of renal clear cell carcinoma needs to be further explored, and experimental basis and research direction for subsequent in-depth research are provided. SUMMARY

[0005] In view of the deficiencies in the prior art, the application provides application of a PELI2 gene as a molecular marker of renal clear cell carcinoma.

[0006] The technical scheme of the application is as follows:

[0007] The application of a reagent for detecting the expression level of a PELI2 gene in the preparation of a renal clear cell carcinoma diagnosis product, wherein the nucleotide sequence of the PELI2 gene is shown as SEQ ID NO. 1.

[0008] According to the application, the reagent for detecting the expression level of the PELI2 gene comprises a real-time fluorescent quantitative PCR detection reagent and a primer pair specific for amplifying PELI2.

[0009] According to the application, the fluorescent quantitative PCR detection reagent comprises fluorescent dye and real-time fluorescent quantitative PCR reaction solution; the real-time fluorescent quantitative PCR reaction solution comprises dNTPs, MgCl2, Taq DNA polymerase and buffer solution; and the fluorescent dye is SYBR Green I.

[0010] According to the application, the sequence of the primer pair for specifically amplifying PELI2 is as follows:

[0011] Upstream primer #1: 5'-GGCTATCAGCTGCAAAGGTC-3',

[0012] Downstream primer #1: 5'-TTCTGTTGATCTGCCCACCT-3'.

[0013] The application of the reagent for detecting the expression level of PELI2 gene in the preparation of a product for evaluating the prognosis of renal clear cell carcinoma, wherein the nucleotide sequence of the PELI2 gene is shown as SEQ ID NO. 1.

[0014] According to the application, the reagent for detecting the expression level of PELI2 gene comprises real-time fluorescent quantitative PCR detection reagent and a primer pair for specifically amplifying PELI2.

[0015] According to the application, the fluorescent quantitative PCR detection reagent comprises fluorescent dye and real-time fluorescent quantitative PCR reaction solution; the real-time fluorescent quantitative PCR reaction solution comprises dNTPs, MgCl2, Taq DNA polymerase and buffer solution; and the fluorescent dye is SYBR Green I.

[0016] According to the application, the sequence of the primer pair for specifically amplifying PELI2 is as follows:

[0017] Upstream primer #1: 5'-GGCTATCAGCTGCAAAGGTC-3',

[0018] Downstream primer #1: 5'-TTCTGTTGATCTGCCCACCT-3'.

[0019] The application of an agonist of PELI2 gene in the preparation of a drug for treating renal clear cell carcinoma, wherein the nucleotide sequence of the PELI2 gene is shown as SEQ ID NO. 1.

[0020] According to the application, the agonist of PELI2 gene comprises but is not limited to nucleic acid molecules, lipids, small-molecule chemicals, antibody drugs, polypeptides or overexpression lentivirus.

[0021] Beneficial effects:

[0022] 1、The present application first found that PELI2 gene is significantly lowly expressed in renal clear cell carcinoma cells, and its expression level is closely related to the prognosis of patients. Specifically, the expression of PELI2 gene in renal clear cell carcinoma cells is statistically different from that in control cells (human renal tubular epithelial cells, HK-2 cells), and the expression of PELI2 gene in renal clear cell carcinoma cells is significantly reduced, and the survival rate of renal cancer patients with high expression of PELI2 gene is significantly higher than that of renal cancer patients with low expression of PELI2 gene. At the same time, the present application verifies the specificity of 72% and the sensitivity of 61% of the PELI2 gene coordinate diagnostic marker when the Youden index is maximum by using the area under the curve (AUC), indicating that PELI2 gene has good diagnostic and prognostic evaluation value for KIRC. The present application designs a primer pair for specifically amplifying PELI2 gene, which can specifically and effectively detect the expression level of PELI2 gene, and is used for preparing a diagnostic and prognostic evaluation kit for KIRC.

[0023] 2、The present application proves by experiments that overexpression of PELI2 gene can more effectively inhibit the proliferation ability of renal clear cell carcinoma cells. On this basis, the present application also designs an overexpression lentivirus of PELI2 gene, which proves that the overexpression lentivirus of PELI2 gene can not only inhibit the proliferation of renal clear cell carcinoma cells, but also can inhibit the expression of proliferation-related gene Ki67 mRNA in human renal clear cell carcinoma cells ACHN and 786O, reduce the expression level of Ki67 mRNA in human renal clear cell carcinoma cells, indicating that overexpression of PELI2 gene can significantly inhibit the DNA replication of human renal clear cell carcinoma cells ACHN and 786O. Therefore, PELI2 gene can be used as a drug target and applied in the research and preparation of renal clear cell carcinoma antitumor drugs. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 is a statistical graph of PELI2 mRNA expression in normal volunteers and renal cancer patient cells.

[0025] Figure 2 is the correlation between PELI2 expression level and prognosis of renal cancer patients.

[0026] Figure 3 is a statistical graph of PELI2 mRNA expression level in human renal clear cell carcinoma cells A498, ACHN, 786O and human renal tubular epithelial cells HK-2.

[0027] Figure 4 is a western blot result graph of PELI2 protein expression in human renal clear cell carcinoma cells A498, ACHN, 786O and human renal tubular epithelial cells HK-2.

[0028] Figure 5 is a receiver operating characteristic curve (ROC) evaluation of the specificity and sensitivity of PELI2 protein expression level in the diagnosis of KIRC.

[0029] Figure 6 is a statistical chart of PELI2 mRNA expression in the PELI2 gene overexpressed renal clear cell carcinoma cell model group and the control group.

[0030] Figure 7 is a western blot result chart of PELI2 protein expression in the PELI2 gene overexpressed renal clear cell carcinoma cell model group and the control group.

[0031] Figure 8 is a statistical chart of cell proliferation inhibition after overexpression of PELI2 gene in human renal clear cell carcinoma cells ACHN and 786O.

[0032] Figure 9 is a result chart (left) and a statistical chart (right) of EDU staining after overexpression of PELI2 gene in human renal clear cell carcinoma cells ACHN and 786O.

[0033] Figure 10 is a statistical chart of Ki67 mRNA expression difference after overexpression of PELI2 gene in human renal clear cell carcinoma cells ACHN and 786O. DETAILED DESCRIPTION

[0034] The technical solutions of the present application will be further described below in combination with examples, but the protection scope of the present application is not limited to this. The reagents and materials involved in the examples are all ordinary commercially available products if not otherwise specified.

[0035] Human renal clear cell carcinoma cells A498, ACHN and 786O and human renal tubular epithelial cells HK-2 were purchased from the National Model and Characteristic Experimental Cell Resource Library / Chinese Academy of Sciences Typical Culture Preservation Committee Cell Library.

[0036] Example 1

[0037] The present inventors found that PELI2 mRNA was significantly lowly expressed in renal clear cell carcinoma patients compared with normal volunteers after analyzing GEO database (GSE53000 dataset, GSE105261 dataset, GSE40435 dataset and GSE53757 dataset). Figure 1 ).

[0038] The Kaplan-Meier Plotter database was used to analyze the impact of PELI2 mRNA expression levels on the prognosis of clear cell renal cell carcinoma. Specific results are as follows: Figure 2 As shown.

[0039] Depend on Figure 2 It can be seen that the overall survival of patients with clear cell renal cell carcinoma in the PELI2 mRNA high expression group was significantly higher than that of patients in the PELI2 mRNA low expression group, which indicates that the expression level of PELI2 mRNA can be used for prognostic assessment of patients with clear cell renal cell carcinoma.

[0040] The specific implementation process is as follows:

[0041] Prognostic information and PELI2 mRNA levels of patients with clear cell renal cell carcinoma were collected using the Kaplan-Meier Plotter database. Based on the median PELI2 mRNA expression level, patients with clear cell renal cell carcinoma were divided into a high-expression group and a low-expression group. The Kaplan-Meier method was used to analyze the difference in overall survival (OS) between the two groups of patients with high and low PELI2 mRNA expression. The statistical significance of the survival difference between the groups was determined by the Log-rank test.

[0042] Therefore, the inventors have for the first time clearly defined the biological role of the PELI2 gene in the progression of clear cell renal cell carcinoma, and believe that the PELI2 gene can be used as a biomarker to prepare products for the diagnosis and prognostic assessment of clear cell renal cell carcinoma, as well as to prepare drugs for targeted therapy of clear cell renal cell carcinoma.

[0043] Example 2

[0044] The study in Example 1 revealed that the PELI2 gene, as shown in SEQ ID NO.1, is abnormally expressed in clear cell renal cancer cells, suggesting that it may serve as a biomarker for the diagnosis, metastasis, and prognostic assessment of clear cell renal cancer.

[0045] The expression level of the PELI2 gene in clear cell renal cancer cells and normal human renal tubular epithelial cells was verified by real-time quantitative PCR.

[0046] 1. The determination of PELI2 gene expression level is carried out as follows:

[0047] (1) Extraction of total RNA: Total RNA was extracted from human clear cell renal cancer cells A498, ACHN, 786O and human renal tubular epithelial cells HK-2 using the TRIzol kit (Invitrogen, USA). The extraction method was strictly performed according to the steps described in the kit instructions. The concentration and purity of RNA were detected using NanoDrop2000.

[0048] (2) Real-time quantitative PCR: The total RNA extracted in step (1) was reverse transcribed into cDNA using a reverse transcription kit (ABI, USA). The specific steps were performed according to the kit instructions. Then, Power SYBR was used. TM Green PCR premix (Thermo Fisher Scientific, 4367659) and the upstream primer #1 / downstream primer #1 provided in this invention were used for real-time PCR, with 18S as an internal control, and 2 -ΔΔCT The method calculates the relative expression level of the PELI2 gene, and the results are as follows: Figure 3 As shown.

[0049] The nucleotide sequences of the primer pairs are as follows:

[0050] PELI2 upstream primer #1: 5'-GGCTATCAGCTGCAAAGGTC-3',

[0051] PELI2 downstream primer #1: 5'-TTCTGTTGATCTGCCCACCT-3'.

[0052] 18S upstream primer: 5'-GCAATTATTCCCCATGAACG-3',

[0053] 18S downstream primer: 5'-GGCCTCACTAAACCATCCAA-3'.

[0054] The reverse transcription PCR system and the real-time quantitative PCR system were established according to the kits.

[0055] Reverse transcription PCR program: 95℃ for 10 minutes; 37℃ for 1 hour, 85℃ for 5 minutes, repeat 2 times; cool to 4℃.

[0056] Real-time quantitative PCR program: 95℃ for 10 minutes; 95℃ denaturation for 30 seconds, 60℃ annealing for 60 seconds, 72℃ extension for 60 seconds, for a total of 40 cycles; 72℃ for 5 minutes.

[0057] Depend on Figure 3It was found that, compared with human renal tubular epithelial cells HK-2, the PELI2 gene was stably and lowly expressed in human renal clear cell carcinoma cells A498, ACHN, and 786O. Furthermore, the expression levels of the PELI2 gene differed significantly among human renal clear cell carcinoma cells A498, ACHN, and 786O and human renal tubular epithelial cells HK-2, indicating that the expression of the PELI2 gene was significantly reduced in renal clear cell carcinoma cells.

[0058] 2. Western blot experiments were performed on human renal clear cell carcinoma cells A498, ACHN, and 786O, and human renal tubular epithelial cells HK-2. The specific procedures are as follows:

[0059] RIPA lysis buffer was added to each group of cells, and the cells were sonicated to disrupt the cell line. The cells were then incubated on ice for 30 min, centrifuged at 12000 rpm and 4°C for 20 min, and the supernatant was collected. SDS loading buffer was added, and the cells were boiled in a metal bath at 95°C for 10 min to denature the protein. The expression level of PELI2 protein was detected by Western blot, using GAPDH as an internal control. The Western blot results are shown below. Figure 4 As shown.

[0060] Depend on Figure 4 It can be seen that the expression level of PELI2 protein in human clear cell renal cancer cells A498, ACHN, and 786O is significantly lower than that in human renal tubular epithelial cells HK-2.

[0061] 3. Thirty-six pairs of surgically resected clear cell renal cell carcinoma (KIRC) specimens were collected. Tumor tissue was taken from the central, non-necrotic area, and adjacent normal tissue was taken from an area more than 5 cm from the tumor margin. All specimens were flash-frozen in liquid nitrogen and stored at -80°C. None of the 36 patients had received preoperative radiotherapy or chemotherapy, and postoperative pathology confirmed KIRC. The collection of these cases was approved by the Medical Ethics Committee of Shandong University, and informed consent was obtained from the patients. The expression level of PELI2 protein was detected in the 36 pairs of KIRC samples. The specific procedure is as follows:

[0062] First, immunohistochemical staining was performed on 36 samples of renal cell carcinoma and 36 samples of adjacent normal tissue. Then, the specificity and sensitivity of PELI2 protein expression level in the diagnosis of KIRC were evaluated using receiver operating characteristic (ROC) curves. The results are as follows: Figure 5 As shown.

[0063] Depend on Figure 5The area under the curve (AUC) was 0.7049 (95% CI: 0.5847–0.8250). When the Youden index was at its maximum, the specificity was 72% and the sensitivity was 61%, indicating that the PELI2 gene has good diagnostic value for KIRC.

[0064] Example 3

[0065] Both the PELI2 gene overexpression lentivirus and the negative control (vector) were generated by coating HEK293T cells. The specific preparation process is as follows:

[0066] (1) HEK293T cells were passaged in advance for transfection and placed in an incubator at 37°C and 5% CO2.

[0067] (2) Observe the cell density before transfection. Transfection can be carried out when the confluence reaches 70-80%.

[0068] (3) Perform lipid-to-complex transfection. The composition of the complex transfected in a 100 mm dish is as follows:

[0069]

[0070] After the lipid-transfer complex was mixed, it was incubated at room temperature for 15 min and then slowly added dropwise to HEK293T cells and cultured in a 37°C, 5% CO2 cell culture incubator.

[0071] (4) Replace with fresh complete culture medium containing 10% fetal bovine serum (FBS) 8 hours after transfection;

[0072] (5) Virus collection: Viral supernatant was collected twice, at 48h and 72h after transfection (after 48h, fresh complete culture medium was replaced).

[0073] At 48 hours, pour the culture medium from the 100mm dish into a 50mL centrifuge tube, ensuring the dish wall does not touch the centrifuge tube opening to prevent bacterial contamination. Then, add 10mL of fresh complete culture medium containing 10% fetal bovine serum (FBS) and incubate stably in a 37°C, 5% CO2 incubator. At 72 hours, directly pour the culture medium from the 100mm dish into a 50mL centrifuge tube, again ensuring the dish wall does not touch the centrifuge tube opening to prevent bacterial contamination.

[0074] According to the product instructions, using polybrene reagent, lentiviruses overexpressing the PELI2 gene and negative controls (vectors) were transfected into human clear cell renal cancer cells ACHN and 786O, respectively, to construct a model group and a control group of clear cell renal cancer cells overexpressing the PELI2 gene. Forty-eight hours after transfection, the expression level of the PELI2 gene in the model group and control group was detected by real-time quantitative PCR. The PCR system and procedure were the same as in Example 2. The results are as follows. Figure 6 As shown.

[0075] Depend on Figure 6 It can be seen that, compared with the control group, the expression level of PELI2 mRNA in human clear cell renal cancer cells ACHN and 786O was significantly increased after transfection with lentivirus overexpressing the PELI2 gene.

[0076] Then, Western blot experiments were performed on the renal clear cell carcinoma model group overexpressing the PELI2 gene and the control group. The specific implementation procedure was the same as in Example 2, and the results are as follows: Figure 7 As shown.

[0077] Depend on Figure 7 It can be seen that, compared with the control group, the expression level of PELI2 protein in human clear cell renal cancer cells ACHN and 786O was significantly increased after transfection with lentivirus overexpressing the PELI2 gene.

[0078] Example 4

[0079] Biological functions of renal clear cell carcinoma cells ACHN and 786O overexpressing the PELI2 gene and control cells constructed in Example 3 were determined.

[0080] 1. CCK8 cell proliferation assay: Renal clear cell carcinoma cells ACHN and 786O overexpressing the PELI2 gene and control cells were cultured at 5 × 10⁻⁶ cells / year. 4 Cells were seeded at a density of 1 / mL in 96-well plates (NEST, China) and cultured. After cell attachment (approximately 2–4 hours), 10 μL of CCK8 solution was added to each well, and the cells were incubated at 37°C for 1 hour. The optical density (OD) at 450 nm was measured using a microplate reader (TECAN, Switzerland). The results are as follows: Figure 8 As shown.

[0081] Depend on Figure 8 It can be seen that overexpression of the PELI2 gene can significantly inhibit the proliferation of human renal clear cell carcinomas ACHN and 786O.

[0082] 2. EdU cell proliferation assay:

[0083] (1) EdU labeling: Renal clear cell carcinoma cells ACHN and 786O overexpressing the PELI2 gene and control cells were labeled with 5 × 10⁻⁶ cells. 4 The cells were seeded at a density of / mL and cultured in the medium. After the cells adhered (about 2-4 hours), medium containing 50μM EdU (reagent A1 diluted 1000) was added to each well and incubated for 2 hours. The medium was then discarded. The cells were washed twice with PBS buffer for 5 minutes each time.

[0084] (2) Cell fixation and permeabilization: Add 500 μL of cell fixation solution (PBS buffer containing 4% paraformaldehyde) to each well of the cells obtained in step (1), fix at 25°C for 30 min, and discard the fixation solution; then add 500 μL of glycine solution with a concentration of 2 mg / mL to each well, place on a decolorizing shaker at room temperature for 5 min to terminate fixation, and discard the solution; then add 500 μL of PBS buffer to each well for washing once, then add 500 μL of PBS buffer containing 0.5% Trinton X-100, place on a decolorizing shaker at room temperature for permeabilization for 10 min, and wash once with PBS buffer;

[0085] (3) Apollo staining: Prepare Apollo staining solution according to Table 1 and use it immediately after preparation; add 500 μL of Apollo staining solution to each well of the cells obtained in step (2), place them on a decolorizing shaker and incubate at room temperature in the dark for 30 min, then discard the staining solution; then add 500 μL of PBS buffer containing 0.5% Triton X-100 to each well and wash 3 times, 10 min each time.

[0086] Table 1

[0087]

[0088] (4) Nuclear staining: The cell nuclei obtained in step (2) were stained and observed and photographed under a fluorescence microscope. The results are as follows: Figure 9 As shown.

[0089] Depend on Figure 9 It can be seen that overexpression of the PELI2 gene can significantly reduce EdU + The proportion of human clear cell renal cancer cells ACHN and 786O indicates that overexpression of the PELI2 gene can significantly inhibit DNA replication in human clear cell renal cancer cells ACHN and 786O.

[0090] 3. Determination of Ki67 mRNA expression level:

[0091] The expression level of Ki67 mRNA in the renal clear cell carcinoma model group overexpressing the PELI2 gene constructed in Example 3 and the control group was detected by real-time quantitative PCR. The PCR system and procedure were the same as in Example 2. The results are as follows.Figure 10 As shown.

[0092] The primer pair sequences for detecting Ki67 mRNA expression levels are as follows:

[0093] Upstream primer #2: 5'-GGATTATACCTGGCCTTCCCC-3';

[0094] Downstream primer #2: 5'-TGACACAACAGGAAGCTGGA-3'.

[0095] Depend on Figure 10 It can be seen that overexpression of the PELI2 gene can significantly inhibit the expression of the proliferation-related gene Ki67 mRNA in human clear cell renal cancer cells ACHN and 786O, and reduce the expression level of Ki67 mRNA in human clear cell renal cancer cells, indicating that overexpression of the PELI2 gene can significantly inhibit DNA replication in human clear cell renal cancer cells ACHN and 786O.

[0096] The embodiments described above are merely preferred implementations of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The application of a reagent for detecting PELI2 gene expression levels in the preparation of diagnostic products for clear cell renal cell carcinoma, characterized in that, The nucleotide sequence of the PELI2 gene is shown in SEQ ID NO.

1.

2. The application as described in claim 1, characterized in that, The reagents for detecting PELI2 gene expression levels include real-time quantitative PCR detection reagents and primer pairs that specifically amplify PELI2.

3. The application as described in claim 2, characterized in that, The reagents include a fluorescent dye for quantitative real-time PCR detection and a real-time quantitative PCR reaction solution. The real-time quantitative PCR reaction solution includes dNTPs, MgCl2, Taq DNA polymerase, and a buffer solution. The fluorescent dye is SYBR Green I.

4. The application as described in claim 2, characterized in that, The sequences of the primer pair for specifically amplifying PELI2 are shown below: Upstream primer #1: 5'-GGCTATCAGCTGCAAAGGTC-3', Downstream primer #1: 5'-TTCTGTTGATCTGCCCACCT-3'.

5. The application of a reagent for detecting PELI2 gene expression levels in the preparation of prognostic assessment products for clear cell renal cell carcinoma, characterized in that... The nucleotide sequence of the PELI2 gene is shown in SEQ ID NO.

1.

6. The application as described in claim 1, characterized in that, The reagents for detecting PELI2 gene expression levels include real-time quantitative PCR detection reagents and primer pairs that specifically amplify PELI2.

7. The application as described in claim 6, characterized in that, The reagents include a fluorescent dye for quantitative real-time PCR detection and a real-time quantitative PCR reaction solution. The real-time quantitative PCR reaction solution includes dNTPs, MgCl2, Taq DNA polymerase, and a buffer solution. The fluorescent dye is SYBR Green I.

8. The application as described in claim 6, characterized in that, The sequences of the primer pair for specifically amplifying PELI2 are shown below: Upstream primer #1: 5'-GGCTATCAGCTGCAAAGGTC-3', Downstream primer #1: 5'-TTCTGTTGATCTGCCCACCT-3'.

9. The application of PELI2 gene agonists in the preparation of drugs for treating clear cell renal cell carcinoma, characterized in that, The nucleotide sequence of the PELI2 gene is shown in SEQ ID NO.

1.

10. The application as described in claim 9, characterized in that, The agonists of the PELI2 gene include, but are not limited to, nucleic acid molecules, lipids, small molecule chemicals, antibody drugs, peptides, or overexpressing lentiviruses.

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