二十碳五烯酸对人肝胆管癌细胞株FRH-0201增殖的影响(1)
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摘要：目的：评价二十碳五烯酸体外对人肝胆管癌细胞株FRH-0201增殖的影响。 方法：在处于指数生长期的FRH-0201细胞株培养剂中添加二十碳五烯酸(EPA)，通过采用MTT法检测EPA体外对FRH-0201细胞的增殖抑制率及IC50，绘制细胞生长曲线以及流式细胞术检测细胞周期来观察EPA对FRH-0201细胞增殖的影响；吖啶橙荧光染色、电镜技术观察FRH-0201细胞凋亡时的形态学特征及超微结构的改变，AnnexinV/PI双染法检测细胞凋亡率。结果：经EPA处理后，FRH-0201细胞增殖受到明显抑制，并且呈明显的量效关系和时效关系。同样条件下正常小鼠成纤维细胞L-929无明显影响。细胞阻滞在G0/G1期。吖啶橙染色细胞内有明显的黄绿色浓染，电镜观察细胞染色质浓缩，EPA 50、70μg/mL作用48h的细胞凋亡率分别为37.3%和62.2%，明显高于正常对照组。结论：EPA体外能通过诱导FRH-0201细胞发生凋亡抑制其增殖。
关键词：二十碳五烯酸(EPA)；人肝胆管癌细胞珠;增值;细胞凋亡率
中图分类号：R285.5文献标识码：A文章编号：1673-7717(2012)03-0556-03
Influence of EPA on Multiplication of Human Cholangiocarcinoma Cell Strain
ZHANG Qing1, WANG Xiangqun1, CHEN Lijuan1, WU Dan2
(1.The First People’s Hospital of Yuhang District, Hangzhou 311100, Zhejiang, China;
2.Department of General Surgery,The Second Affiliated Hospital of Medical School of
Zhejiang University,Hangzhou 310006, Zhejiang, China)
Abstract:Objective: To evaluate the influence of EPA on multiplication of Human cholangiocarcinoma cell strain FRH-0201. Method: Add EPA into exponential growth phase FRH-0201 cell strain. Detect the suppression ratio and IC50 of FRH-0201 cells in vitro with EPA through MTT method. Draw a cell growth curve and FCM detect cell cycle to observe the influence of EPA on mutiplication of FRH-0201 cells. AO fluorescent stain and electron microscope were used to observe the apoptosis of FRH-0201 cells on morphology feature and ultrastructure changes.AnnexinV/PI dubble stain were used to detect the apoptosis rate. Result:After adding EPA, the multiplication of FRH-0201 cells were apparently inhibited in the dose-response and time-response relationship . No response in the mouse fibroblast L-929 within the same condition. The cells were blocked in the G0/G1 phases. Yellow green thick strain was found in the AO stain cells.Chromatin concentration was found in the electron microscopy . The 48h apoptosis rate with 50 and 70μg/mL EPA was 37.3% and 62.2%,apparently higher that of normal control group. Conclusion:EPA will induce the apoptosis of FRH-0201 cells in vitro to inhibit their multiplication.
Key words:eicosapentaenoic acid, EPA; human cholangiocarcinoma cell strain; multiplication; apoptosis rate ......
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