关键词: 缺糖, 热休克蛋白70, Bax, Bcl-2, 荧光免疫法

Abstract: Objective To observe the effect of heat shock protein 70 (Hsp70) on apoptosis of HeLa cells induced by glucose deprivation and investigate the relationship between Hsp70 and the key proteins of apoptosis: Bcl-2 family members. Methods HeLa cells were stably transfected with the plasmid of pcDNA31(+)Hsp70 to establish the Hsp70 overexpressed cell model; Hsp70 normalexpression and overexpression cells cultured with glucose free medium to create stress models. Every detection had three parallel samples. MTT assay was applied to evaluate the cell viability; Hoechst 33258 and Giemsa stain were used to examine the rate of cell apoptosis. RT-PCR was used to determine the expression level of Bax and Bcl-2 and to calculate the ratio of Bax/Bcl-2; Immunofluorescence and immunocytochemistry were applied to examine the conformational change of Bax. Results Cell viability was increased and the rate of cell apoptosis was decreased in Hsp70 overexpression cells cultured with glucose free medium. At the same time, the expression of Bax and Bcl-2 and the conformational change of Bax were also inhibited by Hsp70. Conclusion Hsp7

Key words: Glucose deprivation, Heat shock protein 70, Bax, Bcl-2, Immunofluorescence