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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。
Objective: To evaluate the effect of eicosapentaenoic acid on proliferation of human hepatocellular carcinoma cell line FRH-0201 in vitro. Methods: Eicosapentaenoic acid (EPA) was added into FRH-0201 cell line culture medium in exponential growth stage. The growth inhibition rate and IC50 of EPA on FRH-0201 cells in vitro were determined by MTT assay, and the cell growth curve 0201 cells were observed by flow cytometry to observe the effect of EPA on the proliferation of FRH-0201 cells. Fluorescence staining of acridine orange and electron microscopy were used to observe the morphological changes and ultrastructural changes of FRH-0201 cells. AnnexinV / PI Double staining method was used to detect the apoptosis rate. Results: After treated with EPA, the proliferation of FRH-0201 cells was significantly inhibited, and the dose-response and time-effect relationship were obvious. Under the same conditions, normal mouse fibroblasts L-929 had no significant effect. Cell arrest at G0 / G1 phase. Acridine orange stained cells were significantly yellow-green staining, electron microscopy of cell chromatin condensation, EPA 50,70μg / mL for 48h apoptosis rates were 37.3% and 62.2%, significantly higher than the normal control group. Conclusion: EPA can inhibit the proliferation of FRH-0201 cells in vitro by inducing apoptosis.