Study On The Relationship Between The Level Of Methylation And The Expression Of Tim-3 On The Surface Of Immune Cells And Prostate Cancer

objective: To study the relationship between the methylation level of gene in the tissues of prostate cancer, the expression level of Tim-3 on the surface of immune cells and the clinicopathological features of prostate cancer, to investigate the role of methylation and Tim-3 in the occurrence, invasion and metastasis of prostate cancer.Method: 1) PITX2, WNT5 a, SPARC, EPB41L3, TPM4 and other genes were selected as target genes, methylation levels of 35 cases of benign prostatic hyperplasia were detected by methylation specific PCR method, and then 157 cases of clinical DNA samples were screened, analysis of the relationship between methylation level and clinicopathological features of prostate cancer; 2) The expression of Tim-3 in serum of 35 cases of benign prostatic hyperplasia and 157 cases of prostate cancer were detected by flow cytometry, to analyze the relationship between the expression level of Tim-3 in serum and the clinicopathological features of prostate cancer; the expression levels of Tim-3 in 35 cases of benign prostatic hyperplasia and 157 cases of prostate cancer were detected by immunohistochemical method, statistical analysis of the relationship between Tim-3expression and clinicopathological features of prostate cancer; 3) Western blot method was used to detect the expression level of Tim-3 in prostate cancer LNCa P cells with metastatic potential, analysis of the expression level and invasion and metastasis of prostate cancer; Si RNA interference was used to detect expression of Tim-3 in prostate cancer LNCa P cells, cell proliferation was detected with MTT, the invasion potential of prostate cancer cells was detected by LNCa P Transwell assay. Result: 1) PITX2, WNT5 a,SPARC, EPB41L3, and TPM4 methylation levels were significantly increased in patients with prostate cancer, it suggests that the methylation level of these genes is related to the development of prostate cancer. according to the different PSA levels, the results showedthat the PITX2 methylation rates were 25.10%, 28.64% and 35.82% in PSA<10ng/ml /,10 < PSA < 20ng/ml, PSA > 20ng/ml group, respectively, PITX2 and WNT5 a gene methylation levels in different PSA groups were compared, and the difference was statistically significant(P<0.05); according to the Gleason score is divided into less than7 and more than 8, PITX2 gene methylation rates were 26.31%, 33.16%, there were statistically significant(P<0.05); according to the different stages of TNM group was less than T2 and Tx N1, M1 groups, PITX2 gene methylation rate was 23.13%, 26.64%,38.37%, there were statistically significant differences between three groups(P <0.05).Multivariate analysis revealed that PITX2 methylation levels were most closely related to prostate cancer. 2) The expression levels of CD4+T and CD8+T in peripheral blood of patients with prostate cancer were(4.77+3.56) %,(5.90+4.91) %, respectively,the expression of benign prostatic hyperplasia was(0.96+0.54) %,(0.73+0.62) %, the difference was statistically significant(P<0.05); the expression of Tim-3 in the prostate hyperplasia and prostate cancer group was detected by Tim-3 immunohistochemistry, and the difference was statistically significant(P<0.05).According to the different PSA levels,Tim-3 expression levels were shown to be different by immunohistochemistry staining tissue of prostate cancer, Tim-3 immunohistochemistry score were 3.92+0.75 points,7.63+0.81 points and 10.37+0.35 points in PSA<10ng/ml /, 10<PSA<20ng/ml, PSA>20ng/ml group, respectively, there were statistically significant(P<0.05); according to the Gleason score is divided into less than 7 and more than 8, Tim-3immunohistochemistry score were 11.02±0.96 points, 5.62±0.69 points, there were statistically significant(P<0.05); according to the different stages of TNM group was less than T2 and Tx N1, M1 groups, Tim-3 immunohistochemistry score were 4.27±0.38 points,7.34±0.71 points, 10.31±0.82, and the difference was statistically significant(P<0.05); 3)The proliferation and invasion of human prostate cancer cell was demonstrated by Tim-3interference, The expression levels of Tim-3 in the control group, Tim-3 over expression group, and the Tim-3-si RNA were significant(P<0.05). Conclusion: 1) The methylation level of PITX2 gene in prostate cancer patients was higher than that in benign prostatic hyperplasia, PITX2 methylation level was related to PSA level, Gleason score and TNM stage, There was a close relationship between PITX2 methylation and prostate cancer. 2)The expression level of Tim-3 in serum and tissue of patients with prostate cancer was higher than that of benign prostatic hyperplasia; ccording to the level of PSA, Gleason score and TNM stage, the expression level of Tim-3 was related, the expression level of Tim-3 was closely related to prostate cancer; 3) Downregulation of Tim-3 expression caninhibit the growth of prostate cancer cells, Tim-3 is an important growth factor of prostate cancer; The expression level of Tim-3 was related to the invasion and metastasis of prostate cancer, it can be used as a molecular marker for the invasion and metastasis of prostate cancer; downregulation of Tim-3 inhibits the invasion and migration of prostate cancer cells in vitro.