半枝莲醇提物含药血清对人结肠癌HT-29细胞增殖、凋亡和周期的影响

Ethanol Extracts from Scutellaria Barbata Medicated Serum on Human Colon Carcinoma HT-29 Cell Proliferation,Apoptosis and the Influence of Cycle

目的:研究半枝莲醇提物(EESB,ethanol extract from Scutellaria barbata)含药血清对人结肠癌HT-29细胞株体外诱导其凋亡和周期的变化。方法:体外常规培养人结肠癌HT-29细胞株,制备含药血清,设置空白血清组、含有不同浓度EESB含药血清组、5-FU组,1应用MTT法评价EESB含药血清对HT-29细胞24 h、48 h、72 h的生长抑制率,2Annexin V-PE/7-AAD双染色分析在流式细胞仪上观察EESB含药血清处理HT-29细胞48 h后的细胞凋亡率,3PI法检测细胞周期变化。结果:1与空白组比较,MTT法显示EESB低、中、高剂量含药血清都能抑制HT-29细胞增殖(P<0.05),抑制率与作用时间及血清含药浓度相关,72 h时其达到39%。2同空白组比较,低、中、高剂量含药血清处理HT-29细胞48 h后凋亡率显著高于对照组(P<0.05),同时低、中、高剂量含药血清处理HT-29细胞48 h后,3与空白组比较,G0/G1期细胞明显较对照组增加(P<0.05),提示半枝莲可促进细胞凋亡,并且将细胞生长周期阻滞于G0/G1期,并且S期减少。结论:半枝莲对人结肠癌HT-29细胞有抑制作用,作用机制可能不光与抑制细胞生长有关,还和阻滞细胞生长周期及诱导细胞凋亡有关。

Objective： To investigate the growth inhibition and induced apoptosis effect of medicated serum of ethanol extracts from Gentianella acuta（ EESB） on human colon Carcinoma HT-29 cells. Methods： Conventionally cultivate colon Carcinoma HT-29 cell lines in vitro and prepare medicated serum. Establish the blank groups and the serum containing different concentration EESB drug-containing serum groups and the 5-FU group,respectively. 1The antiproliferative activity of EESB medicated serum on HT-29 cell was evaluated by MTT assays. 2Annexin V-PE /7-AAD double staining assay was used for flow cytometry assay of HT-29 cell apoptosis induced by EESB medicated serum after 48 h.3Determinate the cell cycle by PI staining. Results： 1Compared with the blank group,MTT showed that serum containing EESB with low,medium,high doses inhibited proliferation of HT-29 cells（ P〈0. 05）,which related inhibitive rate to function time and drug concentration of medicated serum,up to 39% in 72 h. 2Compared with the blank group,through low,medium and high dose medicated serum treatment after 48 h HT-29 cell apoptosis rate was significantly higher than that in the control group（ P〈0. 05）. Meanwhile,the low,medium and high doses of medicated serum processing HT-29 cells after 48 h,3compared with the blank group,G0 / G1 phase cell obviously increased than that in the control group（ P〈0. 05）. It showed Scutellaria barbata could promote cell apoptosis,and arrest the cell cycle in G1 / G0 phase,with S phase reduced. Conclusion： Scutellaria barbata has an inhibitory effect on the human colon carcinoma HT-29 cells,and the mechanism may be related to inhibiting the growth of cells,block cell cycle and inducing apoptosis.