摘要
目的 探讨乙醇对肝细胞癌 (简称肝癌 )细胞凋亡的诱导作用 ,并对其相关蛋白的表达进行初步分析。方法 用含 6 %乙醇的培养液培养 6h诱导肝癌细胞系HCC 92 0 4细胞凋亡 ,进行台盼蓝染色、Hoechst 33 2 5 8荧光染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记 (TUNEL)法检测和流式细胞仪DNA含量分析。SDS PAGE电泳分析诱导凋亡前后的细胞蛋白表达的情况。结果 6 %乙醇可使HCC 92 0 4细胞产生明显的凋亡形态学变化。大部分细胞呈台盼蓝拒染 ,并且为TUNEL阳性着色。流式细胞仪分析可见亚二倍体凋亡峰。SDS PAGE电泳显示 ,乙醇诱导凋亡的HCC 92 0 4细胞中有新的分子量约大于 97.4kD的蛋白产生。结论 低剂量乙醇能够明显地诱导HCC 92 0 4肝癌细胞凋亡 。
Objective To explore the apoptosis inducing effect of ethanol on hepatocellular carcinoma(HCC)cells and analyze the expression of its associated proteins primarily.Methods Apoptosis of HCC cell line HCC 9204 cells was induced by culturing the cells for 6h in the culture liquid which contained 6% ethanol.Apoptotic cells were detected with typan blue staining,Hoechst 22 258 fluorescence staining,and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method.DNA contents was analyzed with flow cytometer.The expression of the cellular proteins prior to and after inducing apoptosis was analyzed with SDS PAGE.Results 6% ethanol can make obviously morphological apoptotic changes on HCC 9204 cells.Most of the cells couldn't be stained by trypan blue,and were TUNEL positively stained.Apoptotic sub G1 peak was seen under flow cytometer.SDS PAGE indicated that there were new proteins produced,whose molecular weight was about more than 97.4kD,in apoptotic HCC 9204 cells induced by ethanol.Conclusions Low dose ethanol can induce apoptosis of HCC 9204 cells obviously and produce new high molecular weight proteins.
出处
《中国肿瘤临床与康复》
2001年第2期3-5,共3页
Chinese Journal of Clinical Oncology and Rehabilitation
关键词
乙醇
细胞凋亡
肝细胞癌
蛋白表达
ethanol
apoptosis
hepatocellular carcinoma
protein expression
