Overexpression of hSav1 promotes Mst1-induced apoptosis in HeLa cells.
- Author:
Zhao-Ming LI
1
;
Wei-Cheng LIU
;
Shuo DONG
;
Xue-Lai LUO
;
Xiao-Lan LI
;
De-Ding TAO
;
Jian-Ping GONG
;
Jun-Bo HU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Cell Cycle Proteins; genetics; metabolism; Cytoplasm; metabolism; HeLa Cells; Hepatocyte Growth Factor; genetics; metabolism; Humans; Plasmids; Proto-Oncogene Proteins; genetics; metabolism; Transfection
- From: Chinese Journal of Oncology 2009;31(7):481-484
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo elucidate the effect of hSav1 expression on Mst1-mediated apoptosis in HeLa cells.
METHODSPlasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 micromol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay.
RESULTSPlasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSav1 could be detect from the anti-Mst1 immunoprecipitation complex. The immunofluorescent labeling showed that hSav1 and Mst1 had the same localization in cells. Overexpressed protein hSav1 did not induce a significant cell apoptosis. However, co-expression of hSav1 with Mst1 resulted in a significant increase of apoptosis above the level seen with Mst1 alone (24.5% +/- 2.4% vs. 39.3% +/- 4.0%, P < 0.05).
CONCLUSIONOur findings indicate that hSav1 is a newly identified protein that interacts with Mst1 and augments Mst1-mediated apoptosis.
