视黄酸依赖Fas/RARα融合基因转染HL-60细胞启动的凋亡效应研究

Study on apoptosis of HL-60 cells induced by fusion gene of Fas/RARα

目的 检测视黄酸依赖Fas RARα表达载体转染HL 6 0细胞启动的凋亡效应。方法 成功构建视黄酸依赖Fas RARα表达载体 ,用脂质体转染HL 6 0白血病细胞 ,经四甲基偶氮唑盐 [3 (4,5 dimethylthiaol 2 yl) 2 ,5 diphenyltetrazoliumbromide ,MTT]、流式细胞、DNA梯形带和透射电镜等方法观测视黄酸处理后细胞凋亡效应的发生。结果 以一定剂量的视黄酸处理HL 6 0后 ,细胞增殖受抑 ,呈凋亡性变。结论 视黄酸及其依赖的融合基因表达可协同诱导HL 6

Objective To test the apoptosis of HL-60 cells after transfecting the vector of Fas/RARα fusion gene induced by ATRA. Methods After transfection of fusion gene vector pRARE-Fas/RAR alpha and treatment of RA in tumor cells, the cellular proliferation capacity was observed with MTT assay, the cell cycle distribution and the apoptotic rate was measured with flow cytometry, the DNA ladder was measured. Results The proliferation capacity decreased, the apoptotic rate and the DNA fragmentation rate increased in cells treated by RA. The DNA ladder was detectable in RA-treated cells. The morphological apoptosis changes were observed by transmission electron microscopy. These changes were more obvious in pRARE-Fas/RAR alpha transfected cells than non-transfected cells. Conclusions RA and the product of exogenous fusion gene induced by RA could cooperate in inhibition of cell growth and inducing apoptosis in HL-60 cells.