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目的 :研究 10 羟基喜树碱 (10 HCPT)诱导人肝癌细胞凋亡的作用。方法 :用 10 HCPT以不同终浓度、不同作用时间诱导SMMC 772 1人肝癌细胞。分别以倒置相差显微镜、电镜、荧光显微镜观察其形态学改变 ,提取细胞内小分子DNA进行琼脂糖凝胶电泳 ,并对这些细胞的P5 3蛋白进行免疫组织化学染色。结果 :当 10 HCPT终浓度 >2 .0 μmol·L-1时 ,部分细胞可见典型的凋亡特征 :核固缩、凋亡小体、梯状电泳带。随着 10 HCPT浓度或作用时间增加 ,凋亡率逐渐升高。各组细胞P5 3蛋白免疫组化染色均为阴性。结论 :10 HCPT可诱导SMMC 772 1细胞凋亡 ,其作用效果呈剂量、时间依赖性
Objective : To study the effect of 10 HCPT on the apoptosis of human hepatocellular carcinoma cells. Methods: SMMC 772 1 human hepatoma cells were induced with 10 HCPT at different final concentrations and different time. The morphological changes were observed by inverted phase contrast microscopy, electron microscopy, and fluorescence microscopy. The intracellular small-molecule DNA was extracted and subjected to agarose gel electrophoresis. The P53 protein of these cells was immunohistochemically stained. RESULTS: When the final concentration of 10 HCPT was >2.0 μmol·L-1, typical apoptosis characteristics were observed in some cells: nuclear condensation, apoptotic bodies, and ladder electrophoresis bands. With the increase of 10 HCPT concentration or action time, the apoptosis rate gradually increased. Immunohistochemical staining of P5 3 protein was negative in all groups. Conclusion :10 HCPT can induce apoptosis of SMMC 772 1 cells, and its effect is dose- and time-dependent.