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。
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 uhrastructure changes. AnnexinV/Pl dubble stain were used to detect the apoptosis rate. Result：After adding EPA, the multiplication of FRH -0201 cells were appar- ently inhibited in the dose - response and time - response relationship. No response in the mouse fibroblast L - 929 with- in 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 701xg/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.
出处
《中华中医药学刊》
CAS
2012年第3期556-558,I0005,I0006,共5页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省科技厅基金资助项目(2007C30037)
