The Function Mechanism Of MiR-34a And It's Target Genes In Local Lung Cancer Cells XWLC-05 And YTMLC

Objective: the objective in this study was to investigate the expression of mi R-34 a of the cell lines local lung adenocarcinoma XWLC-05,lung squamous carcinoma YTMLC and other NSCLC,to investigate the effects of mi R-34 a on the proliferation,migration,invasion of the local lung adenocarcinoma XWLC-05,lung squamous carcinoma YTMLC and normal adenocarcinoma H157.And to identify the target protein of mi R-34 a on the NSCLC XWLC-05 and YTMLC cell lines.Methods: Recovery and generation of local lung cancer cell lines of XWLC-05,YTMLC and general lung cancer cell line H157.The three cells XWLC-05,YTMLC and H157 were divided into three groups: blank control group,negative control group and experimental group.p GCMV/EGFP-hsa-mir-34 a mimics transfection experimental group was adopted;the p GCMV/EGFP-NC transfected negative control group was used respectively;No treated cell lines were treated as blank groups.Real-time fluorescence quantitative PCR was used to detective the relative expression of mi R-34 a in lung cancer cells;The migration of cells was detected by scratch test;The invasive ability of each group of samples was tested by Trans-well method.MTT was used to compare the proliferation ability of each cell.Transmission electron microscope experiment method was used to detect the qualitative of apoptosis.Flow cytometry method was used to detect the quantitative of apoptosis in each group.The m RNA expression in each group was detected by real-time fluorescence quantitative PCR.The expression levels of target genes(PTEN,YY-1,CDK-6,C-MET)in each cell were detected by western-blot.Results: Compared with the pulmonary epithelial cell BEAS-2B,Mi R-34 a is low expression in XWLC-05,YTMLC,H157,A549 non-small cell lung cancer cells(P <0.05);In the experimental group(transfection mimics-34a),the grow ability of the experimental group was significantly lower than the blank group and negative control group.The invasion ability is obviously weakened;Under the microscope,the migration ability of the experimental group cells was significantly reduced.The cell apoptosis ratio was significantly increased by flow cytometry.Expression of mi R-34 a in transfection cells was increased in XWLC-05,YTMLC lung cancer cells,the m RNA and protein expression levels of CDK6 were down-regulated(P < 0.05);To the contrary,the m RNA andprotein expression levels of PTEN were up-regulated as well as YY-1,statistically significant difference(P < 0.05),C-MET was not statistically significant.Conclusions: overexpression of mi R-34 a can inhibit the growth and migration of lung cancer cells,and the increase or down-regulation of related target protein suggests mi R-34 a can be used as a molecular target for treatment of lung cancer.mi R-34 a may through the direct or indirect pathway,up or down regulated the expression of protein(PTEN,YY-1,CDK-6)which associated with the important routes of the occurrence and development of cancer,thereby inhibits the behaviors as migration and transfection of cancer cells.Compared with general lung adenocarcinoma cell H157,the cell lines XWLC-05 and YTMLC have no obvious differences in cell function and molecule function.If the Xuanwei lung adenocarcinoma and Gejiu lung squamous carcinoma has obvious differences in biological function,has already developed specific local lung cancer need us further research.