半枝莲提取物诱导结肠癌HT-29细胞凋亡的机制研究  

Mechanism of Apoptosis of Human Colon Cancer HT-29 Cells Induced by Scutellaria barbata D.Don Extract

作　　者：彭军[1] 林久茂[1] 魏丽慧[1] 徐伟[2] 

机构地区：[1]福建中医药大学中西医结合研究院,福建福州350108 [2]福建中医药大学药学院,福建福州350108

出　　处：《福建中医药大学学报》2010年第6期35-38,共4页Journal of Fujian College of Traditional Chinese Medicine

基　　金：福建省自然科学基金(2010J01195);福建省中西医结合老年性疾病重点实验室开放课题(2008J1004)

摘　　要：目的探讨半枝莲提取物在体外诱导结肠癌HT-29细胞株凋亡的作用机制。方法采用人结肠癌细胞HT-29常规体外培养,随机设定对照组和不同浓度半枝莲提取物组,干预24 h。倒置显微镜观察细胞形态变化,MTT法测定不同浓度半枝莲提取物对HT-29细胞增殖的影响,DNA Ladder试剂盒检测药物作用后细胞凋亡情况,AnnexinV-FITC/PI双标记流式细胞术检测细胞凋亡率,JC-1试剂盒检测线粒体膜电位变化,应用比色法分析Caspase-9和Caspase-3的活化情况。结果半枝莲提取物干预HT-29细胞24 h后,HT-29细胞密度明显减少、细胞变小,可明显抑制HT-29细胞增殖,细胞凋亡增加,线粒体膜电位丧失,具有显著地剂量依赖,并出现明显的DNA Lad-der条带;Caspase-9和Caspase-3的活化随着药物浓度的升高而增加。结论半枝莲提取物能通过线粒体通路诱导人结肠癌HT-29细胞凋亡而发挥其抗肿瘤作用。Objective To investigate the mechanism by which the extract of Scutellaria barbata D.Don（ESB） induces apoptosis of human colon carcinoma HT-29 cells.Methods Human colon carcinoma HT-29 cells were cultured in vitro and treated with different concentration of ESB for 24 hours.Cellular morphology of HT-29 cells was observed via phase-contrast microscopy.The cell viability was determined by MTT assay.Cell apoptosis was detected by DNA fragmentation and flow cytometry with Annexin V staining.Mitochondrial membrane potential was examined by flow cytometry with JC-1 staining.Activity of caspase 9 and caspase 3 was examined by colorimetric assay.Results After treated by ESB,the proliferation of HT-29 cells was inhibited,the cell apoptosis increased,and the mitochondrial membrane potential collapsed in a dose dependent manner.The DNA ladder bands appeared and the activity of caspase 9 and caspase 3 increased with the increasing of ESB concentration.Conclusion Extract of ESB induces apoptosis of human colon carcinoma HT-29 cells through mitochondrial pathway,and maybe it is one of the mechanism of the anti-tumor effect of ESB.

关 键 词：半枝莲 结肠癌 HT-29 细胞凋亡 线粒体 

分 类 号：R285.5[医药卫生—中药学]