microrna-195调控lats2对bel-74025-fu细胞凋亡的影响word论文.docxVIP

结论：1、miR-195 能够促进 BEL-7402/5-FU 细胞凋亡。2、miR-195 能够上调 BEL-7402/5-FU 细胞中 LATS2、P53 蛋白的表达，下调 CDK2蛋白表达。3、miR-195 可通过靶向调控 LATS2 促进 BEL-7402/5-FU 细胞凋亡。关键词：MIR-195； LATS2； BEL-7402/5-FU 细 胞；凋亡 2ABSTRACTThe Effection of MiR-195 Regulation of LATS2 Gene on the BEL-7402/5-FU Cells ApoptosisObjective: This study is to investigate the effects of miR-195 and its mechanism on the BEL-7402/5-FU cells apoptosis.Method: MicroRNA microarray analyzed a wide diversity in miRNA expression between BEL-7402 and BEL-7402/5-FU cells. miR-195 or Negative Control, was transient transfected into BEL-7402/5-FU cells. Luciferase assays to identify putative miR -195 targets.Transfection efficiency was tested by fluorescence inverted microscope and Quantitative Real-Time Reverse-Transcription PCR. Cell proliferation of each group was measured by MTT assay. Cell apoptotic percentage was detected by flow cytometry. Western blot was used to detect the protein expression.Result: MicroRNA microarray showed that 2 miRNAs were significant (2- fold) up-regulation (eg.miR-18 and miR-224) and many miRNAs were down-regulation(0.5-fold) (eg. miR-195，miR-18a， miR-122， et al) in the BEL-7402/5-FU comparedwith BEL-7402 cells. MTT results showed that miR-195 transfectants had a lower cell proliferation than untreated cells or negative. Flow cytometry results demonstrated that miR-195s transfected cells had a higher apoptosis rate than untreated cells or negative. Western bolt results showed that the expression level of LATS2 and P53 in miR-195 transfected cells were obviously increased, while the expression level of CDK2 were obviously decreased compared with untreated cells or negative. Dual luciferase activity verified miR-195 regulates its expression by targeting LATS2.Conclusion: 1. miR-195 increased cell spontaneous apoptosis. 2. miR-195 up-regulate LATS2, P53 expressions and down-regulate CDK2 expressions inBEL-7402/5-FU cells. 3. miR-195 could pro-apoptosis of BEL-7402/5-FU cells by3targetti