晚期糖基化终产物对人结肠癌细胞SW-480增殖的影响及其机制研究  

Effects of Advanced Glycosylation End Products on Proliferation of Human Colon Carcinoma Cell Line SW-480 and Its Mechanism

作　　者：周小智[1,2] 薛耀明[1] 关美萍[1] 朱波[1] 

机构地区：[1]南方医科大学南方医院内分泌代谢科,广州510515 [2]吉安市中心人民医院内分泌科

出　　处：《华西医学》2011年第6期837-841,共5页West China Medical Journal

摘　　要：目的观察晚期糖基化终产物(advanced glycosylation end prodrcts,AGE)对人结肠癌细胞株SW-480增殖的影响,并探讨其可能机制。方法不同浓度AGE干预SW-480细胞,噻唑蓝(MTT)法比较各组细胞活力,流式细胞术观察AGE对SW-480细胞周期的影响,蛋白质印迹法观察AGE对SW-480细胞CyclinD1表达的影响,端粒重复序列扩增法(telomeric repeat amplification protocol,TRAP)银染法观察AGE对SW-480细胞端粒酶活性的影响。MTT测细胞活力的检测设置空白对照组、100μg/mL小牛血清白蛋白(bovine serum albumin,BSA)组及50、100、500μg/mL AGE组,其余检测只设置100μg/mL BSA组和100μg/mL AGE组。结果 MTT结果示AGE促进SW-480细胞的增殖,且呈浓度依赖性。100μg/mL BSA组与100μg/mL AGE组72h后的细胞G0/G1期所占百分比分别为56.02%±0.58%、51.93%±1.01%,差异有统计学意义(P<0.05)。蛋白质印迹法示100μg/mL AGE组72h后CyclinD1的表达较100μg/mL BSA组增加,差异有统计学意义(P<0.05)。TRAP银染法检测示100μg/mL AGE干预SW-480细胞72h后可以增加端粒酶活性(P<0.05)。结论 AGE可促进人结肠癌细胞SW-480生长,呈剂量依赖性。其作用机制可能与AGE上调CyclinD1的表达加速G1/S期转换及增加端粒酶活性有关。Objective To observe the effects of advanced glycosylation end products（AGE） on proliferation of SW-480 cells and study the possible mechanism.Methods Various concentrations of AGE were designed to have impact on SW-480 cells.Proliferation of SW-480 cells was assessed by thiazolyl blue tetrazolium bromide（MTT） assay;The impact of AGE on the cell cycle of SW-480 cells was analyzed by flow cytometry（FCM）;the influence of AGE on expression of CyclinD1 was checked by Western blotting;and the impact of AGE on telomerase activity was examined by telomeric repeat amplification proctol（TRAP） sliver staining.For the MTT assay,blank control group,100 μg/mL bovine serum albumin（BSA） group,50,100 and 500 μg/mL AGE groups were designed,while for other examinations,there were only 100 μg/mL BSA group and 100 μg/mL AGE group.Results MTT result showed that AGE increased the proliferation of SW-480 cells in a dose-dependent mode.The proportion of the cells at G0/G1 stage of the 100 μg/mL BSA group and the 100 μg/mL AGE experimental group were（56.02±0.58）% and（51.93±1.01）% respectively after 72 hours,with a significant difference（P0.05）;western blotting showed that the expression of CyclinD1 in the 100 μg/mL AGE group was significantly higher than that in the 100 μg/mL BSA group after 72 hours;TRAP silver staining demonstrated that telomerase activity increased significantly after treated with 100 μg/mL AGE for 72 hours.Conclusions AGE can promote the growth of SW-480 cells in a dose-dependent mode.Its mechanism is mainly by up-regulating the expression of CyclinD1 to shorten G0/G1 and increasing the telomerase activity significantly.

关 键 词：晚期糖基化终产物 SW-480细胞 细胞周期 CYCLIND1 端粒酶活性 

分 类 号：R587.1[医药卫生—内分泌] R735.3[医药卫生—内科学]