The Effection Of MiR-195Regulation Of LATS2Gene On The BEL-7402/5-FU Cells Apoptosis

Objective: This study is to investigate the effects of miR-195and its mechanism on theBEL-7402/5-FU cells apoptosis.Method: MicroRNA microarray analyzed a wide diversity in miRNA expressionbetween BEL-7402and BEL-7402/5-FU cells. miR-195or Negative Control, was transienttransfected into BEL-7402/5-FU cells. Luciferase assays to identify putative miR-195targets.Transfection efficiency was tested by fluorescence inverted microscope andQuantitative Real-Time Reverse-Transcription PCR. Cell proliferation of each group wasmeasured by MTT assay. Cell apoptotic percentage was detected by flow cytometry.Western blot was used to detect the protein expression.Result: MicroRNA microarray showed that2miRNAs were significant (>2-fold)up-regulation (eg.miR-18and 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-7402cells. MTT results showed that miR-195transfectants had a lower cellproliferation than untreated cells or negative. Flow cytometry results demonstratedthat miR-195s transfected cells had a higher apoptosis rate than untreated cells ornegative. Western bolt results showed that the expression level of LATS2and P53inmiR-195transfected cells were obviously increased, while the expression level ofCDK2were obviously decreased compared with untreated cells or negative. Dualluciferase activity verified miR-195regulates its expression by targeting LATS2.Conclusion:1. miR-195increased cell spontaneous apoptosis.2. miR-195up-regulate LATS2, P53expressions and down-regulate CDK2expressions inBEL-7402/5-FU cells.3. miR-195could pro-apoptosis of BEL-7402/5-FU cells by targetting LATS2.