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目的体外培养人肝癌细胞株 HepG-2,BEL-7402,观察测定应用不同浓度三氧化二砷后多种指标的变化,从多个角度探讨三氧化二砷的抗肿瘤作用及其机制.方法应用倒置相差显微镜、电子显微镜、透射电镜、扫描电镜、流式细胞仪、细胞免疫组化法,分别对不同浓度加药组及对照组 HepG-2,BEL-7402细胞的存活,形态学改变,细胞 DNA含量的分布以及 Bcl-2,Bax 蛋白的表达进行了观察和测定.结果 0.5,1,2μmol/L As_2O_3均能抑制人肝癌细胞株细胞的生长增殖.流式细胞仪分析显示,加药组在 G_1期细胞前均出现亚二倍体峰,且 G_0/G_1期细胞减少,S 期细胞增多,电镜下,对照组细胞核质比大、核大、核膜有明显切迹,0.5μmol/L As_2O_3组细胞核质比减小、核变圆、胞质内出现分化良好的细胞器,0.5,1,2μmol/L As_2O_3组均可见细胞膜完整、核固缩、凋亡小体形成.细胞免疫组化法测定 Bcl-2,Bax 蛋白的表达以及两者之间的比率均有变化.结论三氧化二砷不仅抑制人肝癌细胞增殖,而且诱导细胞凋亡.诱导肝癌细胞凋亡与 Bcl-2,Bax 蛋白的表达改变有关.
Objective To culture human hepatocellular carcinoma cell lines HepG-2 and BEL-7402 in vitro, observe and determine the changes of various indexes after applying different concentrations of arsenic trioxide, and explore the anti-tumor effects and mechanisms of arsenic trioxide from multiple perspectives. Methods Inverted phase contrast microscope, electron microscope , Transmission Electron Microscopy, Scanning Electron Microscopy, Flow Cytometry, Cellular Immunohistochemistry, Survival, morphological changes, distribution of cellular DNA, and Bcl in HepG-2 and BEL-7402 cells at different concentrations -2, Bax protein expression was observed and measured. Results 0.5,1,2μmol/L As_2O_3 can inhibit the growth and proliferation of human hepatocellular carcinoma cell lines. Flow cytometry analysis showed that the drug group was in the G_1 phase cells before Subdiploid peaks appeared, and cells in G 0 /G 1 phase decreased, and cells in S phase increased. Under electron microscopy, the control group had a large nuclear-to-cytoplasmic ratio, a large nucleus and a clear notch in the nuclear membrane, and a 0.5 μmol/L As 2 O 3 group had a cytoplasmic nuclear reduction ratio. Small cell, round nuclear, and well-differentiated organelles were observed in the cytoplasm. The cell membrane integrity, nuclear condensation, and apoptotic bodies were observed in 0.5, 1, and 2 μmol/L As 2 O 3 groups. Bcl-2 and Bax were measured by immunohistochemistry. Protein expression And the ratio between them has changed. Conclusions Arsenic trioxide not only inhibits the proliferation of human hepatoma cells, but also induces apoptosis. The induction of apoptosis of hepatoma cells is related to the changes of Bcl-2 and Bax protein expression.