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目的观察C23过表达对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响。方法先构建pRSET-C23重组质粒并将其导入SaOS-2细胞;再将细胞随机分为四组:对照组、单纯用CDP组、转染空载体后再用CDP组与转染C23重组质粒后再用CDP组;用MTT法检测各组SaOS-2细胞增殖率,用Western Blot和RT-PCR方法检测各组C23、Bcl-2与Bax蛋白和mRNA的表达情况;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 pRSET-C23重组质粒构建成功;转染C23重组质粒后再用CDP组的增殖率与单纯用CDP组比较,升高了约16%(P<0.01),C23与Bcl-2的蛋白和mRNA表达均增高,但Bax表达则降低(P<0.01);转染C23重组质粒后再用CDP组与单纯用CDP组比较,细胞凋亡核百分率、细胞凋亡率均降低,差异有统计学意义(P<0.01)。结论 C23过表达能抑制CDP诱导的SaOS-2细胞增殖抑制和凋亡。
Objective To observe the effect of overexpression of C23 on proliferation and apoptosis of human osteosarcoma cell line SaOS-2 induced by CDP. Methods The recombinant plasmid pRSET-C23 was constructed and introduced into SaOS-2 cells. The cells were randomly divided into four groups: the control group, the pure CDP group, the vector transfected with empty vector, the CDP group and the C23 recombinant plasmid The proliferation of SaOS-2 cells was detected by MTT assay. The protein and mRNA expression of C23, Bcl-2 and Bax in each group were detected by Western Blot and RT-PCR. The percentage of apoptotic nuclei and apoptotic rate in each group were detected by cytometry. Results The recombinant plasmid pRSET-C23 was constructed successfully. The proliferation rate of transfected with C23 recombinant plasmid was about 16% (P <0.01) higher than that of CDP group alone. The protein and mRNA of C23 and Bcl-2 (P <0.01). Compared with CDP group, the percentage of apoptotic nuclei and apoptosis rate of C23 recombinant plasmids transfected with C23 recombinant plasmid were lower than those of CDP alone group, the difference was statistically significant (P <0.01). Conclusion C23 overexpression can inhibit CDP-induced proliferation and apoptosis of SaOS-2 cells.