| Objective:To observed the effects on cell proliferation, migration, apoptosis and analysis the expression of target gene EZH2and COX-2protein in cancer cells when up-regulated the expression of miR-101And then to explore the role of miR-101in cervical cancer cells.Methods:The synthetic mimics of miR-101(up-regulated group) and inhibitor (re-duced group) transfected into cervical cancer cell lines (Hela, Siha) by LipofectamineTM2000,with the esophageal squamous cell carcinoma cell lines Eca109as a control group. Application qRT-PCR to detect the expression of miR-101in each group of cells, MTT assay was used to detect cell proliferation activity, By flow cytometry to detect cell apoptosis, Scratch experiments to tested with cell migration capability, Immunohistochemical method to detect the expression of EZH2and COX-2protein.Results:The qRT-PCR results of two cell lines:In the up-regulated group, the expression levels of miR-101is higher than untransfected group; in the lower group, the expression levels of miR-101is lower than untransfected group. The results of MTT:After96h, the up-regulated group, cell proliferation is reversed; in the down-regulated group the results is increased. Flow cytometry results:In the untransfected group, Hela, Siha apoptosis rates were12.2%,8.5%, up-regulated group, the apoptosis rates were97.6%,76.6%,, down-regulated group, apoptosis rate is decrease, Siha was21.6%. Scratch Results:In the up-regulated group, Hela cells, scratches distance(42.65um±2um) is lower than untransfected group (181.38um±2um). Siha, scratches distance (36.63um±2um) is lower than untransfected group (184.12um±2um). Immunohistochemical results:up-regulate group, EZH2, COX-2protein positive rate was lower than untransfected group.while, the down-regulated group, the positive rates of the two proteins was increased compared with untransfected group. These results were statistically significant differences (P<0.05).Conclusions:Over-expression miR-101is able to inhibits cancer cell proliferation inhibits cancer cell migration, promote cancer cell apoptosis,.miR-101plays an important role in cervical carcinogenesis process. EZH2, COX-2is one of the target genes of miR-101,both of which showed a negative regulator of relationships. |
