蓝舌病毒HbC_3对人肾癌细胞的感染杀伤作用

目的 观察蓝舌病毒湖北株(BTV-HbC_3)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-HbC_3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(MTT)比色法检测细胞存活率,RNA凝胶电泳检测BTV-HbC_3在ACHN细胞内的增殖,DNA ladder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-HbC_3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,1 MOI的BTV-HbC_3感染ACHN细胞24、36、48 h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1 MOI的BTV-HbC_3感染ACHN细胞24、36、48 h后可见低亚二倍体峰,比例分别为13.7%、18.5%和24.2%.结论 BTV-HbC_3能有效地杀伤人肾癌ACHN细胞.

The infectivity of bluetongue virus strain HbC_3 to human renal cell carcinoma cells

Objective To study the characteristics of human renal cell carcinoma ACHN cells in-duced by bluetongue virus strain HbC_3 (BTV-HbC_3 ) and the action mechanism. Methods BTV-HbC_3 was used to infect the tumor cells. DNA ladder, and flow cytometry were used to detect the apoptosis of tumor cells induced by BTV-HbC_3. Agarose gel electrophoresis was used to detect the RNA of tumor cells induced by BTV-HbC_3. The changes of endoplasmic reticulum and nuclei treated with BTV-HbC_3 were fur-ther examined by electron microscopy. Results Cytopathie effect (CPE) was observed. The electron mi-croscopy examination showed characteristics of necrocytosis, and lots of BTV-HbC_3 appeared in cytoplasm. The 24-, 36-and 48-h survival rate of ACHN cells infected with BTV at 10 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-HbC_3 was decreased sharply and the result of DNA ladder showed untypical apoptosis. Mild sub-G_1 was observed by flow cytometry. The apopto-sis 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-HbC_3. These data provide the basis for the possibility of BTV-HbC_3 being the oncolytic virotherapy.