摘要
目的 探讨雷公藤内酯醇（TPL）对人胃癌细胞SGC-7901增殖、凋亡及上皮间质转化（EMT）的影响。方法常规培养SGC-7901细胞达对数生长后,采用终浓度为12.5、25、50、100 nmol/L TPL处理SGC-7901细胞,应用四甲基偶氮唑盐比色法测定对照组及不同浓度TPL处理24、48、72和96 h的细胞增殖情况,采用流式细胞术检测不同浓度TPL处理48 h的凋亡率和细胞周期分布情况,实时荧光定量聚合酶链反应检测不同浓度TPL处理48 h的促EMT转录因子Snail1、Twist的表达情况,Western blotting检测各组处理48 h后的EMT相关标记分子（E-cadherin、N-cadherin及Vimentin）水平。结果 TPL在12.5~100 nmol/L的范围内对SGC-7901细胞有增殖抑制作用,且增殖抑制率呈剂量和时间依赖性,且除24 h 12.5 nmol/L外,其余浓度及作用时间的增殖抑制率均高于对照组（P〈0.05）;与对照组相比,TPL处理48 h后的早晚期凋亡率升高、G0/G1期细胞比例及E-cadherin水平均升高,S期细胞比例、N-cadherin和Vimentin蛋白水平及Snail1和Twist mRNA水平均降低,差异均有统计学意义（P〈0.05）,且呈浓度依赖性。结论 TPL对胃癌细胞SGC-7901有毒性作用,不仅抑制其增殖且诱导凋亡及细胞周期G0/G1期阻滞,同时可抑制其EMT过程。
Objective To investigate the effects of triptolide （TPL） on the proliferation, apoptosis and epithelial mesenchy- real transition （EMT） of human gastric cancer cell line SGC-7901. Methods The SGC-7901 cells of logarithmic growth were treated with different final concentrations of TPL at 12. 5, 25, 50 and 100 nmol/L. The MTI＂ method was employed to detect the cell prolifera- tion of cells treated with different concentrations of TPL at 24, 48, 72 and 96 h. The apoptotic rate and cell cycle distribution of cells exposure to different concentrations of TPL at 48 h were measured by flow cytometry. Real time fluorescence quantitative polymerase chain reaction was used to detect the expression of Snaill and Twist in different groups treated with TPL at 48 h. Western blotting was used to detect the protein levels of EMT-related markers, including E-cadherin, N-cadherin and Vimentin at 48 h. Results TPL in the range of 12. 5-100 nmol/L exhibited the inhibitory effects on SGC-7901 cells in a dose-and time-dependent manner. In addition to cells exposure to 12. 5 nmol/L for 24 h, the proliferation inhibition rates of the rest concentrations and time were higher than the control group （P〈0. 05 ）. Compared with the control group, there were elevated early/late apoptotic rates, proportion of cells at G0/G1 phase and protein level of E-cadherin but decreased proportion of cells at S phase, protein levels of N-cadherin and Vimentin and mRNA lev- els of Snaill and Twist in TPL-treated cells （P〈0. 05）. Conclusion TPL can inhibit the proliferation and induce apoptosis and Go/ G1 phase arrest of gastric cancer cells SGC-7901. Besides, it has the effect on the inhibition of EMT process.
出处
《临床肿瘤学杂志》
CAS
2016年第10期883-888,共6页
Chinese Clinical Oncology
关键词
雷公藤内酯醇
胃癌
增殖
凋亡
上皮间质转化
Triptolide
Gastric cancer
Proliferation
Apoptosis
Epithelial mesenchymal transition
