摘要
目的建立肝癌细胞凋亡模型，为进一步研究细胞凋亡分子机理打下基础。方法用抗癌药阿霉素诱导培养的人肝癌细胞株ＨＣＣ－９２０４肝癌细胞发生凋亡，用四唑盐比色试验（ＭＴＴ法）检测。结果研究发现，０．５μｇ／ｍｌ放线菌素Ｄ可明显抑制２０μｍｏｌ／Ｌ阿霉素对肝癌细胞凋亡的诱导作用。用２０μｍｏｌ／Ｌ阿霉素作用肝癌细胞３小时，换培养液继续培养１２小时后，肝癌细胞ＤＮＡ结构断裂，电泳呈梯状带型。肝癌细胞形态学改变出现膜小泡和凋亡小体形成等凋亡细胞特征。用流式细胞仪测定，出现典型的凋亡峰，凋亡细胞占７２．１％。用２０μｍｏｌ／Ｌ阿霉素诱导肝癌细胞凋亡后，细胞周期Ｇ１期、Ｓ期和Ｇ２期分别为７５．９％，２４．１％和０％，肝癌细胞活细胞率为３５．５％。结论用抗癌药阿霉素成功地诱导人肝癌细胞株ＨＣＣ－９２０４肝癌细胞发生凋亡，该细胞凋亡模型的建立，将有助于进一步探讨肝癌细胞凋亡的分子机制。
Objective Establishment of cell apoptotic model of human hepatocellular carcinoma. Methods Human cultured hepatocellular carcinoma cell line HCC 9204 was exposed in vitro to different concentrations of adriamycin for vairous lengths of time. Cell apoptosis was ascertained by cell morphology under transmission electron microscope, DNA electrophoresis and flow cytometry. Results When HCC 9204 cells were treated with adriamycin at 20 μmol/L for 3 hr, cell apoptosis occurred. Typical apoptotic bodies appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptotic peak was also shown by flow cytometry. Apoptotic cells accounted for 72.1% of the cell population. Cells in G 1, S and G 2 phase of cell cycle were 75.9%,24.1% and 0%, respectively. Only 35.5% of adriamycin treated cells remained alive. Conclusion The apoptosis of human cultured hepatocellular carcinoma cells, HCC 9204 cell line, was successfully induced by adriamycin. Establishment of this model will help further explore the molecular mechanism of hepatoma cell apoptosis.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
1997年第4期260-263,共4页
Chinese Journal of Oncology
基金
国家自然科学基金
关键词
肝肿瘤
细胞凋亡
阿霉素
模型
Liver neoplasms Apoptosis Carcinoma, hepatocellular Adriamycin Cell cycle Cell line
