Expression of Mortalin in human hepatoma-derived cell lines and its role in epithelial-mesenchymal transition
10.3760/cma.j.issn.1007-8118.2014.01.0014
- VernacularTitle:热休克蛋白75在肝癌细胞株中的表达及其在上皮间质转化中的作用
- Author:
Jing CHEN
;
Wenbin LIU
;
Weidong JIA
;
Geliang XU
;
Jinliang MA
;
Jiansheng LI
- Publication Type:Journal Article
- Keywords:
Hepatocellular carcinoma;
Mortalin;
Epithelial-mesenchymal transition;
Vimentin
- From:
Chinese Journal of Hepatobiliary Surgery
2014;20(1):51-55
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the expression of Mortalin in human hepatoma-derived cell lines and explore its effect on epithelial-mesenchymal transition in hepatocellular carcinoma (HCC) cell lines.Methods Six HCC cell lines and 1 normal liver cell line (L02) were chosen.The expression of Mortalin was detected using Western blot and real-time quantitative PCR (qPCR).The endogenous gene expression of Mortalin was inhibited by RNA interference (shRNA).Cell viability was detected using MTT assay and flow cytometry.The expression of Mortalin,E-cadherin and Vimentin were detected by Western blot and qPCR.The experiment was divided into three groups; blank,control,and shRNA.Results Mortalin was detected in Hep3B,MHCC97H,HepG2,and HCCLM3,but not in MHCC97L and L02.After 24 h transfection,GFP fluorescence showed that plasmid Mortalin shRNA was successfully transfected into MHCC97H cells.MTT assay indicated that cytotoxicity was 0％,2.5％,and 3.5％ in the blank,control,and shRNA group respectively.Similarly,flow cytometric showed that early apoptosis rates were 0.8％,4.5％,and 9.2％ in the blank,control,and shRNA group respectively.These results indicated that transfection did not cause severe cell damage.After 48 h of interference,Western blot and qPCR analysis showed that shRNA significantly inhibited the expression of Mortalin.Moreover,cells were collected after 24 h,48 h,72 h and 96 h of interference and analyzed for the relationship between Mortalin,E-cadherin and Vimentin by Western blot and qPCR.It was found that decreased expression of Mortalin was accompanied by elevated E-cadherin expression and reduced Vimentin expression.Conclusion Overexpression of Mortalin correlated with the metastatic phenotype of HCC cells and could promote epithelial-mesenchymal transition.
