摘要
目的观察不同浓度雷公藤甲素作用于人肝癌细胞株HepG2细胞，检测其体外生长增殖抑制率、细胞凋亡率及细胞内Ca^2＋浓度的变化，探讨其诱导肝癌细胞凋亡的机制。方法应用不同浓度的雷公藤甲素（12．5、25、50、100μ∥L）处理HepG2细胞，用噻唑蓝（MTT）比色法测定不同作用时间细胞增长抑制率；流式细胞仪（FcM）膜联蛋白V／碘化丙锭（AnnexinV／PI）双染色法检测细胞凋亡率的变化；采用荧光探针技术（nuo一3）和激光共聚焦显微镜，测定细胞内^2＋浓度的变化。结果（1）雷公藤甲素（12．5、25．0、50．0、100．0μg／L）对人肝癌细胞HepG2增殖有抑制作用（P〈O．05），抑制率与药物浓度、作用时间呈正相关；（2）雷公藤甲素（12．5、25．O、50．O、100μg∥L）能够诱导人肝癌细胞HepG2发生凋亡，随药物作用时间的延长、药物浓度的增高，细胞凋亡率逐渐增高；（3）作用时间为24、48h时，细胞内ca。’荧光强度与药物浓度、作用时间呈正相关，与对照组比较差异有统计学意义（P〈0．01），而作用时间至72h时，荧光强度逐渐减弱甚至恢复至正常水平（P〉0．05）。结论雷公藤甲素能够抑制人肝癌细胞HepG2的生长增殖，其效应呈时间、剂量依赖性；且能诱导HepG2凋亡，其诱导凋亡分子机制可能与细胞内^2＋浓度变化有关。
Objective To observe the inhibitory effects of different concentrations of triptolide on in vitro growth, proliferation, apoptosis and intracellular Ca2＋ of human liver cancer cell line HepG2. Methods In vitro, HepG2 cells were treated with triptolide in various concentrations respectively （ 12. 5, 25.0, 50.0 and 100.0 txg/L）, and examined at 24, 48 and 72 h. The inhibitory effect on cell growth was assayed by using MTY method. Cell apoptosis rate was detected by using flow cytometry （FCM） with an- nexin V/propidium iodide （Annexin V/PI） double stainings. The intracellular Ca2^＋ was determined by u- sing a laser scanning confocal microscope. Results （1） After treatment with triptolide （12. 5, 25.0, 50. 0, and 100. 0 tLg/L）, the cell proliferation was inhibited in time- and dose-dependent manners. The ap- optosis morphologic changes of HepG2 cells treated with triptolide （12. 5, 25.0, 50. 0, and 100. 0μg/L） were observed through AnnexinV-PI staining. AnnexinV-PI staining showed that triptolide could induce HepG2 cell apoptosis and the apoptosis ratio was time- and dose-dependent （ P 〈 0. 05 or P 〈 0. 01 ）. The concentration of intracellular Ca2^＋ in HepG2 cells treated with triptolide （ 12. 5, 25.0, 50.0 and 100. 0 ixg/L） at 24 and 48 h was statistically different from that in the control group cells （P 〈0. 01 ）, bat at 72 h, the concentration of intracellular Ca^2＋ was gradually reduced and even showed no stastieally differ- ent from that in control group cells （P 〉 0.05 ）. Conclusion Triptolide could inhibit the growth of human liver cancer cell line HepG2 in a dose- and time-dependent manner. The molecular mechanism of triptolie inducing apoptosis of HepG2 cells may be related to the increased concentration of intracellular Ca^2＋ in HenG2 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第3期502-504,共3页
Chinese Journal of Experimental Surgery
