靛玉红甲肟对肝癌HepG2细胞周期和凋亡的影响

Effect of Indirubin-3’-monoxime on Proliferation and Apoptosis of HepG2 cells

目的探讨靛玉红甲肟对肝癌HepG2细胞增值和凋亡的影响。方法分别用0、10、20、40μmol/L靛玉红甲肟作用于HepG2细胞24、48、72h,采用MTT法检测细胞抑制率;用0、10、20、40μmol/L靛玉红甲肟作用于HepG2细胞24h后收集样品,采用流式细胞术法检测细胞周期及其凋亡率。结果 MTT法测定靛玉红甲肟对肝癌HepG2细胞的增殖具有抑制作用,且表现出剂量和时间的依赖性,随着浓度升高时间加长,抑制明显(P<0.01)。流式细胞术测定也表明靛玉红甲肟对肝癌HepG2细胞有诱导凋亡的作用,凋亡随着靛玉红甲肟浓度的增大不断地增加。细胞形态学方面也可观察出细胞随着靛玉红甲肟剂量和时间的增大而逐渐出现越来越多的凋亡,细胞核染色质深染,聚集于核膜下呈新月形,细胞膜突起呈小泡样,小泡脱落形成含核小体片段的凋亡小体。结论靛玉红甲肟确实可以诱导HepG2细胞凋亡,但靛玉红甲肟对肝癌细胞作用机制还不明确,需要作进一步的深入研究。

Objective To discuss the effect of indirubin-3＇-monoxime on proliferation and apoptosis of HepG2 cells. Method HepG2 cells were treated with 0, 10, 20, 40μmol/L indirubin-3＇-monoxime for 24, 48, 72hours, then the apoptosis rate was measured by methabenzthiazuron （MTT） assay, also, it was measured by flow cytometry. Use the coloring agent DAPI and PI to dye the HepG2 cells which had been treated, then observe the cell morphology under the fluorescence microscope. Results Indirubin-3＇-monoxime inhibited growth of HepG2 cells in a dose-dependent and time-dependent. The apoptosis rate increased after the treatment by indurubin-3＇-monoxime at 24 hours, this apoptosis are differences between different dose group. In cell morphology, we can observe that more and more cells apoptosis along with the increasing of a dose and .time of indurubin-3＇-monoxime.Chromatin of cell nucleus concentrating and anachromasis, gathering in nuclear membranes as crescent-shaped. Cell membranes raised as bubble. The bubble will fall off and formed apoptotic body which contain fragment of nucleosome. Conclusion Indirubin oxime can indeed induce apoptosis in HepG2 cells, but Indirubin oxime on hepatoma cell mechanism is not clear, the need for further in-depth study.