叶黄素对人结肠癌HT29细胞增殖的抑制及其机制  

Mechanisms underlying suppressive effect of lutein on cell proliferation in human colon cancer cell line HT29

作　　者：付蕾[1] 陈晓哲[1] 张慧娟[1] 张源渊[1] 王凌飞[1] 徐少博[1] 张玉杰[1] 王明臣[2] 

机构地区：[1]郑州大学药学院,河南省郑州市450000 [2]郑州大学基础医学院生物化学教研室,河南省郑州市450000

出　　处：《世界华人消化杂志》2013年第13期1239-1244,共6页World Chinese Journal of Digestology

基　　金：河南省医学科技攻关基金资助项目;No.20060078~~

摘　　要：目的:研究叶黄素对结肠癌HT29细胞的增殖抑制作用及可能的机制.方法:用不同浓度的叶黄素(20、40、80、160 m g/L)和空白对照组(0 m g/L)干预处理培养的HT29细胞24、48、72 h,采用SRB法检测其对该细胞的增殖抑制作用;流式细胞仪检测细胞周期;Hoechst33342/PI荧光染色法检测细胞凋亡;Western blot检测磷酸化ERK(phosphorylation of ERK,p-ERK)、磷酸化p38(phosphorylation of p38,p-p38)蛋白表达水平的变化.结果:叶黄素能抑制HT29细胞增殖,且有明显的剂量和时间依赖性.流式细胞仪检测细胞周期结果显示,叶黄素(80 mg/L)干预处理HT29细胞48 h后,G0/G1期细胞由58.67%增加至63.23%,且随药物浓度增加,G0/G1期细胞显著增加,当叶黄素浓度为160 mg/L时,G0/G1期细胞增加至70.81%,表明叶黄素可将HT29细胞阻滞在G0/G1期;Hoechst33342/PI荧光染色法检测细胞凋亡结果显示,叶黄素可诱导HT29细胞凋亡;Western blot检测结果显示叶黄素可下调p-ERK、上调p-p38蛋白的表达,有浓度依赖性(P<0.01).结论:叶黄素可显著抑制HT29细胞的增殖并诱导其凋亡,使细胞周期阻滞在G0/G1期;下调p-ERK蛋白、上调p-p38蛋白的表达可能是其诱导细胞凋亡的重要机制.AIM：To study the anti-proliferative effect of lutein on human colon cancer HT29 cells and to explore the possible mechanisms involved.METHODS：HT29 cells were treated with different concentrations of lutein（20,40,80,160 mg/L） for 24,48 or 72 h.After treatment,cell proliferation was detected by SRB assay,cell cycle progression was analyzed by flow cytometry,cell apoptosis was detected by fluorescence microscopy and Hoechst 33342/PI staining,and the levels of p-ERK and p-p38 proteins were determined by Western blot.RESULTS：Lutein treatment inhibited the proliferation of HT29 cells in a dose-and time-dependent manner.After treatment of HT29 cells with lutein（80 mg/L） for 48 h,the percentage of cells at G0/G1 phase cells increased from 58.67% to 63.23%,and with the increase in drug concentration,the percentage of cells at G0/G1 phase increased significantly.When the lutein concentration was 160 mg/L,the percentage of cells at G 0/G1 phase increased to 70.81%,indicating that lutein arrested HT29 cells in G0/G1 phase.Fluorescence microscopy and Hoechst 33342/PI staining suggested that lutein induced HT29 cell apoptosis in a dose-dependent manner.Western blot analysis showed that lutein down-regulated the expression of p-ERK and up-regulated the expression of p-p38 protein in dose-dependent manner（both P〈0.01）.CONCLUSION：Lutein could significantly inhibit the proliferation of HT29 cells,induce apoptosis,and arrest cell cycle in G0/G1 phase.Down-regulation of p-ERK protein expression and up-regulation of p-p38 protein expression may be important mechanisms responsible for the apoptosis-inducing effect of lutein.

关 键 词：叶黄素 结肠癌 增殖 P-ERK P-P38 

分 类 号：R735.35[医药卫生—肿瘤]