Abstract
Objective To investigate the Vero cell apoptosis caused by the protein ICP27 of herpes simplex virus type Ⅱ and its possible way involved in apoptosis.Methods The successfully constructed pcDNA3.0-ICP27 were transiently transfected into Vero cells.The viability of Vero cells transfected with pcDNA3.0-ICP27 was detected by MTT assay.The effect of ICP27 on Veto apoptosis was detected by Gimesa stain and flow cytometry.Real time fluorescent quantitative PCR was carried out to detect the change of Bax and Bcl-2 mRNAs,and the Caspase-3 was detect by Caspase-3 kits to investigate the pathway of apoptosis induced by ICP27.Results MTT assay showed that the viability of Vero cells transfected with pcDNA3.0-ICP27 was obviously lower than the Vero cell transfected with no plasmid or transfected with pcDNA3.0.Vero cells transfected with pcDNA3.0-ICP27 became round and the disrupted nucleus was observed by Giemsa stain.Flow cytometry showed that Vero cells transfected with pcDNA3.0-ICP27 had higher apoptosis rate.The expression of Bax increased,while the expression of Bcl-2 decreased in Vero cells transfected with pcDNA3.0-ICP27.Compared with the normal Vero cells and the Vero cells transfected with pcDNA3.0,Bax/Bcl-2 significantly increased in the Vero cells transfected with pcDNA3.0-ICP27.Conclusions HSV-2 ICP27 might induce apoptosis by changing Bax/Bcl-2 ratio in the course of reactivation from latent. Key words: HSV-2 ; ICP27 ; Eukaryotic expression ; Apoptosis
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