核仁素表达下调对顺铂诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响

目的:观察核仁素(C23)表达下调对顺铂(DDP)诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响。方法:将细胞随机分为5组:对照组、DDP组、S+DDP组(转染C23正义寡核苷酸后再用DDP)、AS+DDP(转染C23反义寡核苷酸后再用DDP)与R+DDP(转染随机寡核苷酸后再用DDP);用MTT法检测各组细胞增殖率;用Western印迹和RT-PCR检测各组C23,Bcl-2和Bax蛋白与mRNA的表达;用DAPI染色方法和流式细胞术检测各组细胞凋亡核百分率、细胞凋亡率。结果:C23反义寡核苷酸显著抑制了C23蛋白和mRNA的表达,与对照组比较,差异有统计学意义(P<0.01);AS+DDP组与DDP组比较,细胞增殖率下降约18%(P<0.01),C23与Bcl-2蛋白和mRNA表达均降低(P<0.01),但Bax表达增高(P<0.01);细胞凋亡核百分率与细胞凋亡率均增高(P<0.01)。结论:C23表达下调能促进DDP诱导的SaOS-2细胞增殖抑制和凋亡。

Effect of C23 down-regulation on proliferation inhibition and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells

Objective: To investigate the effect of C23 down-regulation on proliferation inhibition and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells. Methods: SaOS-2 cells were randomly divided in to 5 groups: a control group, a DDP group, an S+DDP group, an AS+DDP group, and an R+DDP group. Cell growth suppression of each group was analyzed by MTT assay. RT-PCR and Western blot were used to detect the expression of C23, Bcl-2 and Bax mRNA and protein in each group. Morphologic character of apoptosis was evaluated by DAPI staining. The percentage of apoptotic cells was analyzed by flowcytometer assay. Results: C23 antisense oligonucleotides remarkably inhibited the expression of C23 mRNA and protein in SaOS-2 cells (P<0.01). The proliferation of SaOS-2 cells was remarkably inhibited in the AS+DDP group compared with the DDP group (P<0.01). The expression of C23, Bcl-2 mRNA and protein was weaker and that of Bax mRNA and protein was stronger in the AS+DDP group compared with the DDP group (P<0.01). Cisplatin-mediated apoptosis neclei in SaOS-2 cells and the rate of apoptotic cells were higher in the AS+DDP group than those of the DDP group (P<0.01). Conclusion: Down-regulation of C23 can contribute to cisplatin-mediated proliferation inhibition and apoptosis in SaOS-2 cells.