hSav1蛋白高表达对Mst1介导的细胞凋亡的影响

Objective To elucidate the effect of hSavl expression on Mstl-mediated apoptosis in HeLa cells. Methods Plasmids 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 μmol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay. Results Plasmids 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 hSavl could be detect from the anti-Mstl immunoprecipitation complex. The immunofluorescent labeling showed that hSavl and Mstl had the same localization in cells. Overexpressed protein hSavl did not induce a significant cell apoptosis. However, co-expression of hSavl with Mstl resulted in a significant increase of apoptosis above the level seen with Mstl alone (24. 5% ± 2.4% vs. 39.3% ± 4.0%, P < 0. 05). Conclusion Our findings indicate that hSavl is a newly identified protein that interacts with Mst1 and a, augments Mst1-mediated apoptosis.

Overexpression of hSav1 promotes Mst1-indneed apoptosis in HeLa cells