蓝舌病毒HbC3对人肾癌细胞的感染杀伤作用

The infectivity of bluetongue virus strain HbC3 to human renal cell carcinoma cells

目的观察蓝舌病毒湖北株（BTV—HbC3）对人肾癌ACHN细胞的感染特性，探讨其杀伤肾癌细胞作用及其机制。方法BTV—HbC3感染肾癌ACHN细胞，观察细胞病变效应及显微电镜变化，噻唑蓝（MTY）比色法检测细胞存活率，RNA凝胶电泳检测BTV—HbC3在ACHN细胞内的增殖．DNAladder法检测感染后凋亡并用流式细胞仪检测感染后细胞凋亡指数。结果BTV—HbC3感染细胞后有明显的细胞病变效应，细胞胞质内出现病毒颗粒，细胞肿胀破裂，1MOI的BTV—HbC3感染ACHN细胞24、36、48h的存活率分别为68．14±5．13、57．14±2．68、42．21±5．63。RNA电泳出现L1—L3、M4-M6和S7-S10特征性病毒dsRNA，DNA ladder显示非特异凋亡表现，流式细胞仪检测1MOI的BTV—HbC3感染ACHN细胞24、36、48h后可见低亚二倍体峰，比例分别为13．7％、18．5％和24．2％。结论BTV-HbC3能有效地杀伤人肾癌ACHN细胞。

Objective To study the characteristics of human renal cell carcinoma ACHN cells induced by bluetongue virus strain HbC3 （ BTV-HbC3 ） and the action mechanism. Methods BTV-HbC3 was used to infect the tumor cells. DNA ladder, and flow cytometry were used to detect the apoptosis of tumor cells induced by BTV-HbC3. Agarose gel electrophoresis was used to detect the RNA of tumor cells induced by BTV-HbC3. The changes of endoplasmic reticulum and nuclei treated with BTV-HbC3 were further examined by electron microscopy. Results Cytopathic effect （CPE） was observed. The electron microscopy examination showed characteristics of necrocytosis, and lots of BTV-HbC3 appeared in cytoplasm. The 24-, 36-and 48-h survival rate of ACHN cells infected with BTV at l0 MOI was 68. 14 ± 5. 13, 57.14±2.68,42.21 ± 5.63 respectively. The detection of RNA showed the characteristic dsRNA bands L1-L3, M4-M6 and S7-S10. The viability of the cells induced by BTV-HbC3 was decreased sharply and the result of DNA ladder showed untypical apoptosis. Mild sub-G1 was observed by flow cytometry. The apoptosis rate of the ACHN cells which were induced by BTV at 24, 36 and 48 h was 13.7%, 18.5% and 24. 2% respectively. Conclusion ACHN cells were sensitive to BTV-HbC3. These data provide the basis for the possibility of BTV-HbC3 being the oncolytic virotherapy.