反义RNA抑制c-myc表达增强骨肉瘤细胞对顺铂的敏感性

Antisense RNA of c-myc gene enhances sensitivity of osteosarcoma cells to cisplatininduced apoptosis

目的通过对骨肉瘤细胞MG-63和U2OS c-myc基因干扰,初步探讨顺铂对c-myc低表达骨肉瘤细胞凋亡的影响。方法以腺病毒为载体,构建表达反义c-myc的重组腺病毒(Ad-Asc-myc),并在体外感染骨肉瘤细胞MG-63和U2OS,与不同浓度及不同时间点的顺铂作用后,采用Western blot方法检测干扰水平;瑞氏染色和MTT检测顺铂对骨肉瘤细胞MG-63和U2OS凋亡作用。结果构建包装的腺病毒Ad-Asc-myc体外感染MG-63和U2OS 48h后,可明显降低cmyc蛋白表达;随着顺铂浓度和处理时间增加可增强对细胞增殖的抑制。结论抑制c-myc的表达可以增强骨肉瘤对化疗的敏感性。

Objective To investigate whether down-regulation of c-myc gene with antisense RNA can increase the sensitivity of osteosarcoma cells MG-63 and U2 OS to cisplatin-induced apoptosis.Methods The recombinant adenovirus（Ad-Asc-myc） encoding antisense RNA targeting c-myc was constructed and used to transfect osteosarcoma cells MG-63 and U2 OS.The expression of c-myc gene was analyzed by quantitative realtime PCR and Western blotting.The change of the sensitivity to cisplain-induced apoptosis was assayed by MTT assay.The shape of osteosarcoma cells was observed by Wright ’s staining.Results Ad-Asc-myc downregulated the expression of c-myc gene after transfection of MG-63 and U2 OS cells for 48 h,which resulted in an increase of cell proliferation inhibition induced by cisplatin in a dose-and time-dependent manner.Conclusion The inhibition of c-myc gene expression increases the chemotherapy sensitivity of osteosarcoma.