五株蓝舌病毒对宫颈癌HeLa细胞作用特性的比较研究

目的 体外研究五株蓝舌病毒(bluetongue virus)致HeLa细胞的作用机制.方法 透射电镜观察感染病毒后细胞超微结构的变化;MTT法和流式细胞仪分别检测病毒对细胞的生长和凋亡的影响;检测凋亡细胞片段化DNA和检测病毒诱导细胞后Caspase-3、-8、-9活性.结果 大量细胞发生病变,出现凋亡和溶解,胞质内可见病毒包涵体及未装配成熟的亚病毒颗粒;五株病毒以浓度依赖方式抑制细胞增殖,且五株病毒的效果有差异;流式细胞仪检测可见各个凋亡阶段的凋亡细胞,并且五株蓝舌病毒感染HeLa细胞的凋亡百分比不一样;DNA-Ladder典型;Caspase-3、-8、-9活性有不同程度的升高.结论 蓝舌病毒能够有效地诱导体外培养的人宫颈癌HeLa细胞进入凋亡,五株蓝舌病毒之间差异明显.

Comparative analysis of targeted oncolysis of five strain bluetongue virus

Objective To study the death mode and mechanism of HeLa cancer cell induced by five strain bluetongue virus(BTV). Methods Transmission electron microscope(TEM) was introduced to study changes of ultrastructure. Growth and apoptosis of HeLa cell infected with bluetongue virus were detected with MTT assay and flow cytometry. DNA fragmentation and the activity of caspase-3, -8, -9 were determined by colorimetric assay. Results Many HeLa cells which infected with BTV were observed apoptosis and lyse, and in the plasma were found many viral inclubodies and subviral particles without outer layer proteins. BTV could inhibit HeLa cell proliferation moderately and different serotypes of virus had different effect. Various stages of apoptotic cells were found by flow cytometry and the percentage of apoptosis caused by five strain bluetongue virus were not the same. DNA-Ladder was typical. Caspase-3,-8 ,-9 activity were increased by varying degrees. Conclusion BTV could infect in HeLa cell efficiently and induce it to apoptosis in vitro, then different serotypes of virus have different effect.