联合应用罗格列酮与维甲酸对HCT-15细胞增殖和凋亡的影响  

The proliferation and apoptosis by PPARr agonist Rosiglitazone (ROS) combined with all-trans retinoic acid(ATRA) on the human colorectal cancer cell line HCT-15

作　　者：郭仁乐[1] 苗瑞政[1] 徐涛[1] 刘立青[1] 王猛[1] 姜言明[1] 

机构地区：[1]山东大学附属省立医院胃肠外科,山东济南250021

出　　处：《中国现代普通外科进展》2010年第6期421-425,429,共6页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:探讨联合应用罗格列酮(ROS)与维甲酸(ATRA)对人结直肠癌细胞株HCT-15细胞增殖和凋亡的影响。方法:实验分为ROS单药组(ROS浓度分别为6.25、12.5、25、50μmol/L)、ATRA单药组(ATRA浓度为2μmol/L)、ROS与ATRA联合用药组(ROS以上各浓度+2μmol/L ATRA)。在分别或联合应用不同浓度ROS和ATRA干预人结直肠癌细胞株HCT-15不同时间后,MTT法检测药物对细胞生长的抑制作用;流式细胞仪检测细胞发生凋亡和细胞生长周期的变化;免疫细胞化学法检测干预前后细胞COX-2、MMP-7和TIMP-1表达蛋白水平的变化。结果:单独应用ROS可抑制HCT-15细胞的增殖,并呈浓度-时间依赖性,与ATRA联合应用则有明显的协同作用(q>1.15)。流式细胞术结果显示:单独应用ROS和ATRA可导致G0/G1期细胞比例上升,而S期比例下降明显,诱导HCT-15细胞凋亡;而联合应用对细胞G1期的阻滞作用更强(P<0.05)。免疫细胞化学结果显示:COX-2、MMP-7和TIMP-1在对照组人结直肠癌细胞HCT-15中阳性表达率分别为66.79%、73.21%、64.08%,联合应用后3种蛋白的表达率为19.33%,20.58%,13.13%,二者差异有统计学意义(P<0.01)。结论:ROS和ATRA可降低HCT-15细胞中COX-2、MMP-7和TIMP-1的表达,使细胞周期阻滞于G1期,诱导细胞凋亡,从而发挥抑制细胞增殖的作用。Objective： To investigate the proliferation and apoptosis mechanism of PPARr ago nist Rosiglitazone（ROS）combined with all- trans retinoic acid（ATRA）on the human colorectal cancercell line HCT- 15. Methods： HCT- 15 cells were affected by different concentrations of ROS or （and） ATRA for different time. MTT assay was used to examine the growth inhibition. Flow cytometry （FCM）was used to examine the apoptosis rate and percentage of cells in each stage of the cellcycle. Expression of COX- 2, MMP- 7, TIMP- 1 were examined by SP immunocytochemical. Results： MTT assay results showed that the growth of HCT- 15 cells treated with ROS alone was inhibited, and had a concentration and time dependence. Combined with the application of ATRA the inhibition was significantly synergy（q1.15）. FCM results showed that the application of ROS or ATRA alone could cause G0 / G1 phase ratio to rise, but S phase decline effectively, and induce apoptosis （P0.05）;while the application of ROS and ATRA caused G1 cell cycle to arrest more strongly.Immunocytochemistry showed that HCT- 15 cells expressed COX- 2, MMP- 7 and TIMP- 1. Thepositive expression in the control group was 66.79% , 73.21% , 64.08% respectively, while the positive expression dropped to only 19.33% ,20.58% ,13.13% after combined application of ROS and ATRA. Conclusion： ROS and ATRA have an effect on inhibiting cell proliferation in the human colorectal cancer cell line HCT- 15 by reducing the expression of COX- 2, MMP- 7, TIMP- 1 and causingcell cycle arrested at G1 stage and inducing apoptosis.

关 键 词：罗格列酮 维甲酸 HCT-15细胞 细胞增殖 细胞凋亡 

分 类 号：R735.3[医药卫生—肿瘤]