MiR495 Gene Promoter Methylation On PRL-3 Expression In Gastric Cancer Cell

AIM: To investigate in gastric cancer peritoneal invasion and metastasis the endogenous mi RNA molecular targeted regulatio n of PRL-3 of mi R-495 gene upstream Cp G island methylation in gastric cancer cel s of PRL-3 gene expression.METHODS: The normal gastric mucosa cell lines and gastric cancer cell lines were maintained in the incubator at 37℃ in an atmosphere of 5%CO2 in medium supplemented with 10% fetal bovine serum. PRL-3 protein levels were detected by Western Bloting. Expression level of mi R-495 was determined by real-time PCR. Methylation specific PCR(MSP) and sodium bisulfite sequencing method(bisulfite genomic sequencing, BSP) detected mi R-495 gene promoter methylation levels. Moreover, Transwell cambers experiment were used to test the ability of invasion and metastasis of tumor cells. Then, apply different concentrations(0、1、5、10μmol/L)of specific DN A methyltransferase inhibitor 5-aza-2’-dc to deal with gastric cancer cell lines, using the above method determination of PRL-3 protein levels, mi R-495 expression, mi R-495 gene promoter methylation level and the change of the ability of invasion and metastasis of tumor cel s.RESULTS: Gastric cancer cells with DNA methyltransferase inhibitors- nitrogen impurity cytidine 5-Aza-2’-deoxycytidine to methylation process, mi R-495 expression were significantly increased, and significantly inhibited the PRL-3 protein and m RNA expression, tranwell affect migration experiments show that after treatment in demethylation, gastric cancer cell invasion and migration ability were significantly lower.CONCLUSION: In gastric cancer peritoneal invasion and metastasis the endogenous mi RNA molecular targeted regulation of PRL-3 of mi R-495 gene upstream Cp G island methylation down-regulation of mi R-495 expression,and lead to up regulate theexpression of PRL-3 gene in gastric cancer peritoneal metastasis.