The Role Of MRI1 Gene In Prostate Cancer Metastasis

With the change of environment,high fat diet and aging population,the incidence of prostate cancer in China increased from 6.53/10 million in 2003 to 9.92/10 million in 2009,an increase of 51.91%,showing an upward trend year by year.At present,radical prostatectomy is regarded as the gold standard for local treatment of localized prostate cancer.There are also endocrine treatments,chemotherapy and radiation.Early prostate cancer has no obvious clinical symptoms,the diagnosis of prostate cancer rely mainly on prostate specific antigen PSA(prostate-specific antigen PSA)and digital rectal examination(digital rectal examination,DRE)examination method.The 5 year survival rate of patients with localized prostate cancer is almost 100%,but the survival rate can be reduced by 29% when distant metastasis occurs.Once it happens,it not only affects the prognosis,but also affects the quality of life of the patients.Tumor metastasis is one of the important biological characteristics of malignant tumor.In recent years,the recurrence and metastasis of malignant tumors affect the prognosis and treatment of the patients.Most patients are not died of primary tumors,but died of cancer metastasis formed after tumor metastasis.With the deepening of research on people and tumor metastasis related genes,found that the Ras(Ras superfamily Ras)and Myc gene family(myelocytomatosis viral oncogene,Myc)and mutant P53 gene(protein 53,P53)the abnormal expression and tumor metastasis associated.In addition,studies have shown that if a lot of cancer gene transfection into the appropriate recipient cells can induce the formation of metastatic phenotype.In the Rho(Ras homolog gene family,Rho)of the family gradually,called Rho A(Ras homolog gene family on member,A,Rho A)invasion of the media "(Mediator of Rho A-dependent invasion,MRI1)has not been identified genes,also known as the MRI1 gene.It has often been considered as a hypothetical protein that has never been characterized in human characteristics data.It is named for its response to Rho A based on the induction and promote feature dependent invasion and cell motility of Rho A cells,and provides a new approach for the regulation of tyrosine phosphorylation of Rho A FAK(Focal,Adhesion,Kinase,FAK).It has dual functions,one is its catalytic activity,and the other is the involvement of cancer cells as an effector signal.Previous studies have shown that MRI1 gene may be associated with metastasis of melanoma,ovarian cancer,breast cancer and other tumors.However,the relationship between prostate cancer and prostate cancer has been reported less.Objective:In this study,we detected the expression of MRI1 gene and its corresponding protein in prostate cancer cells and prostate cancer cells after silencing MRI1 gene.The effect of silencing MRI gene on the migration of prostate cancer cells was observed by cell transfection,and the role of MRI1 gene in prostate cancer cell metastasis was investigated.Material and method:1.The object of study: PC3 human prostate cancer cell line(protate cancer cell 3,PC3),ATCC(American type culture purchased from collection),cells containing 10% fetal bovine serum RPMI1640 medium at 37 DEG C and 5% CO2 and saturated humidity conditions.2.Experimental grouping: experimental groups were divided into three groups,the first group was the control group(CTL group)for prostate cancer cells without any treatment,the second groups as negative control group(NC group)to join the empty lentiviral vector,third as experimental group(sh-1,sh-2,sh-3,sh-4,sh-5)silencing of MRI1 gene.3.Methods: using lentiviral vector to specifically target silencing MRI1 gene expression,and using real-time quantitative PCR and Western blot to detect the expression of MRI1 gene and protein in prostate cancer cells.The migration ability of prostate cancer cells and silent MRI1 gene in prostate cancer cells was detected by scratch test.4.Data processing: application of statistical software SPSS 19,the relative expression of MRI1 gene in prostate cancer cells after treatment in different values of the independent samples T test was used for analysis and comparison of ability of migration of prostate cells,by analysis of variance.Test level alpha ?=0.05.Result:1.Real time quantitative PCR results showed that MRI1 gene was expressed in prostate cancer cells,the MRI1 gene in the experimental group was more effective than the CTL group,and the expression of MRI1 gene decreased significantly(P<0.05).2.Western blot results showed that the reference protein bands of Vinculin order,all samples show that the concentration of protein is consistent.Compared with the protein expression in the two control group,the protein expression in the experimental group was significantly decreased.3.Scratch experiment results show that the experimental group after 8h scratch width the final scratch width is significantly higher than the control group,we will crawl into the experimental cell width of quantitative analysis,found that the migration of prostate cancer cells after MRI1 gene silencing was significantly reduced(P<0.05).Conclusion:The over expression of MRI1 may be closely related to the metastasis of prostate cancer.Silencing MRI1 gene can effectively reduce the metastasis of prostate cancer.