阿霉素诱导人肝癌细胞凋亡模型的建立

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。

Establishment of an apoptotic model induced by adriamycin in human hepatocellular carcinoma

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 various 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 mumol/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 G1, S and G2 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.