10-羟基喜树碱诱导人肝癌细胞SMMC-772l凋亡及其机制的初步研究

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-721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P53蛋白进行免疫组织化学染色。结果当10-HCP终浓度>2.0μmol·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P53蛋白免疫组化染色均为阴性。结论10-HCPT可诱导SMMCC-7721细胞凋亡，其作用效果呈剂量、时间依赖性。

Induction of Apoptosis by 10-Hydroxycamptothecin in SMMC-7721 Human Hepatoma Cells

Objective To investigate the effect of 10-hydroxycamptothecin(10-HCPT) on the induction of apeptosis in human hepatoma cells. Methods SMMC-7721 human hepatoma cells were treated with 10-HCPT in different concentration or for different time in vitro. Their morphological changes were detected by light microscopy, electron microscopy and fluorescence microscopy. In addition, low molecular-weight DNA was isolated fr om those cells, and then subjected to electtophoresis through a agarose gel. Furthermore, P53 protein in those cells was investigated by immunohistochemistry. Results When concentration of 10-HCPT was higher than 2.0 μmol· L- 1, the hallmarks of apoptosis were detected in some cells ( condensed chromatin,apoptosis bodies,“ladder” pattern of intemucleosomal DNA cleavage). The percentage of apoptotic cells rose with increasing concentration of 10-HCPT or prolonging exposure time. Immunohistochemistry staining for p53 protein was negative in both treated and untreated cells. Conclusion 10-HCPT can induce apoptosis in SMMC-7721 human hepatoma cells. The efficiency of 10-HCPT depended on both drug concentration and exposure time.