摘要
目的观察汉防己甲素诱导人胃癌BGC-823细胞自噬与凋亡的作用,并探讨两者发生以及相互关联的分子机制。方法用不同浓度的汉防己甲素作用于人胃癌BGC-823细胞,用四甲基偶氮唑盐比色法(MTT法)检测细胞增殖率;流式细胞术检测细胞凋亡情况;Western-blot法检测各组细胞Bcl-2和自噬标记蛋白LC3、Beclin1的表达,MDC自噬特异性染料荧光染色观察自噬泡的聚集。结果 TET对BGC-823细胞的生长有显著抑制作用,呈明显的时间、剂量依赖性;TET可诱导BGC-823细胞发生凋亡;TET可诱导BGC-823细胞发生自噬,自噬作用在8、10μg/mL TET给药组达到高峰,后随着浓度的增加逐渐下降;在TET给药前用3-甲基腺嘌呤(3-MA)阻断自噬后,低浓度(8μg/mL)TET诱导的凋亡率明显升高;高浓度(20μg/mL)TET诱导的凋亡率差别无统计学意义。结论 TET能明显抑制人胃癌BGC-823细胞的生长,并诱导其发生凋亡和自噬。TET诱导的自噬作为保护性机制,可发挥拮抗凋亡的作用。
Objective To investigate the effects of tetrandrine( TET)-induced death of human gastric cancer cell line BGC-823,and to explore potential mechanisms involved between autophagy and apoptosis. Methods After treatment with different concentrations of tetrandrine,the growth of BGC-823 cells was detected by MTT method. The flow cytometry was performed to detect the apoptosis. The expression of BCL-2,LC3 and Beclin 1 was detected by Western blotting. MDC autophagy specific fluorescence staining was used to evaluate the autophagy in the BGC-823 cells treated with tetrandrine. Results The growth of BGC-823 cells was significantly inhibited in a dose-and time-dependent manner with tetrandrine treatment. TET could induce autophagy in the BGC-823 cells. TET could induce apoptosis in the BGC-823 cells,which reached the peak after being treated with TET at the concentrations of 8 μg / mL and 10 μg / mL and then gradually decreased. With 3-MA pre-treatment to inhibit autophagy,apoptosis rate induced by low concentration of TET( 8 μg / mL)was significantly increased,and there was no statistically significant difference in high concentration( 20 μg / ml) TET-induced apoptosis rate compared with that of the control group. Conclusions TET could significantly inhibit the growth of human gastric cancer cell line BGC-823,and induce the autophagy and apoptosis. In the process of TET-induced death of BGC-823 cells,autophagic mechanism might play a protective role in resisting the apoptosis.
出处
《山东医药》
CAS
2013年第30期1-5,共5页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81070423)
