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目的研究Bax对HCC-9204细胞系凋亡和对阿霉素敏感性的影响。方法采用脂质体转染法将Bax基因转入HCC-9204细胞中。免疫组织化学分析Bax在HCC-9204细胞中的表达,TUNEL及细胞周期分析细胞凋亡,MTT实验判断肿瘤细胞的活力。结果免疫组化结果显示转染Bax基因的HCC-9204/Bax细胞的Bax表达显著增加。TUNEL检测显示,HCC-9204/Bax细胞的凋亡指数在ZnSO4诱导后24、48h明显增加(3·6±5·3vs27·2±10·5,t=63·45,P<0·05;4·2±4·1vs32·3±8·6,t=93·27;P<0·05),流式细胞仪分析显示,诱导后48h出现显著的“亚二倍体峰”。MTT实验显示Bax基因的过表达能够降低阿霉素处理后的细胞活力,呈时间依赖性。结论Bax基因不仅可以诱导HCC-9204细胞的凋亡,而且能够增加HCC-9204细胞对阿霉素杀伤作用的敏感性。
Objective To investigate the effect of Bax on the apoptosis and the sensitivity to doxorubicin in HCC-9204 cell line. Methods The Bax gene was transfected into HCC-9204 cells by lipofection. Immunohistochemical analysis of Bax expression in HCC-9204 cells, TUNEL and cell cycle analysis of apoptosis, MTT assay to determine the activity of tumor cells. Results The results of immunohistochemistry showed that Bax expression was significantly increased in HCC-9204 / Bax cells transfected with Bax gene. TUNEL assay showed that the apoptotic index of HCC-9204 / Bax cells was significantly increased at 24 and 48 h after ZnSO4 induction (3.6 ± 5.3 vs27.2 ± 10.5, t = 63.45, P <0.05) 4 · 2 ± 4 · 1vs32 · 3 ± 8 · 6, t = 93 · 27; P <0 · 05). Flow cytometry analysis showed significant “subdiploid peak” 48h after induction. MTT experiments showed that Bax gene overexpression can reduce the doxorubicin-treated cell viability in a time-dependent manner. Conclusion Bax gene can not only induce the apoptosis of HCC-9204 cells, but also increase the sensitivity of HCC-9204 cells to doxorubicin killing.