Effects of MAGEA3 inhibition by shRNA on apoptosis in human hepatocellular cancer cells
- VernacularTitle:RNA干扰MAGEA3对人肝癌细胞凋亡的影响
- Author:
Wenyu LI
;
Ailin GUO
;
Suqing YANG
;
Shaowei HU
;
Jianming WEN
- Publication Type:Journal Article
- Keywords:
RNA interference;
Melanoma associated antigen A3;
Liver neoplasms;
Apoptosis
- From:
Chinese Journal of Pathophysiology
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the inhibitory effect of vector-based RNA interference(RNAi) on the expression of melanoma associated antigen A3(MAGEA3) protein in hepatocellular carcinoma cells and on apotposis of hepatocellular carcinoma cells.METHODS: A vector for transcribing specific small hairpin RNA(shRNA) targeting MAGEA3 gene was constructed,introduced into hepatocellular carcinoma MEL-ED1 cells by Lipofectamine 2000.The MAGEA3 protein and mRNA expression levels of MEL-ED1 cells were detected by Western blotting and RT-PCR, respectively.The cell apoptosis was studied by DNA fragmentation,electron microscopy,TUNEL assay,and annexin V/PI staining.RESULTS: The vector of RNA interference was successfully constructed and MAGEA3 expression was descreased significantly in MEL-ED1 cells.After the shRNA expression vector was transfected into the MEL-ED1 cells,the expression of MAGEA3 gene was inhibited significantly(by 90%).DNA fragmentation,electron microscopy and TUNEL assay showed classic apoptosis characters in the MEL-ED1 cells transfected with pSilencer-MAGEA3 plasmid with an apoptosis rate of 21.41% ?1.98%,significantly higher than those in the negative control group transfected with pSilencer-neo and in the non-transfected group(both P
