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目的评价柯萨奇病毒B3CVB3致HeLa细胞死亡的方式及分子机制。方法用CVB3作用于HeLa细胞,在不同时间收集细胞,通过相差显微镜、电子显微镜、流式细胞仪以及分子生物学手段,检测HeLa细胞的病变及caspase-3基因mRNA和蛋白质的表达。结果CVB3作用于HeLa细胞后,细胞很快发生变性和坏死,24h后较多细胞凋亡;caspase基因在病毒作用早期即被活化,表现在caspase-3mRNA在病毒作用后6h内,迅速增高达峰值。在24h内,又降至接近病毒作用前的水平。caspase-3蛋白表达在42h内逐渐增高。结论CVB3可诱导HeLa细胞发生坏死和凋亡两种反应,坏死早于凋亡,细胞凋亡与caspase-3的表达密切相关。
Objective To evaluate the molecular mechanism of HeLa cell death induced by Coxsackievirus B3CVB3. Methods HeLa cells were treated with CVB3 and cells were collected at different times. The pathological changes of HeLa cells and the expression of caspase-3 mRNA and protein were detected by phase-contrast microscopy, electron microscopy, flow cytometry and molecular biology. Results After treated with CVB3, the cells were rapidly degenerated and necrotic, and more cells were apoptosised after 24 hours. The caspase gene was activated in the early stage of the virus. The expression of caspase-3 mRNA rapidly increased up to peak within 6 hours after virus exposure . Within 24 h, it dropped to the level before pre-virus effect. The expression of caspase-3 gradually increased within 42h. Conclusions CVB3 can induce necrosis and apoptosis in HeLa cells. The necrosis is earlier than apoptosis, and apoptosis is closely related to the expression of caspase-3.