摘要
目的研究全长脂联素(Acrp30)在人乳腺癌MCF-7细胞增殖、迁移、自噬、凋亡中的作用,并探讨自噬与凋亡的关系。方法体外培养MCF-7细胞,分为对照组(未加入Acrp30)、加药组(25、50、100、200 ng/ml Acrp30),四亚基偶氮唑盐(MTT)比色法检测各组细胞增殖情况。选取100 ng/ml Acrp30浓度组(Acrp组)作用MCF-7细胞,进行培养,倒置显微镜观察细胞形态、贴壁情况;划痕抑制试验了解细胞迁移能力;Western blot法评价细胞自噬水平;流式细胞仪检测Acrp组及3-甲基腺嘌呤(3-MA)预处理组不同时间细胞的总凋亡率。结果 1 MTT结果显示:与对照组相比,50、100 ng/ml组72 h,200 ng/ml组48、72 h可显著抑制MCF-7细胞增殖(P<0.05);2划痕抑制实验结果显示:与对照组相比,Acrp30组作用48、72 h,迁移距离显著减小(P<0.01);3 Western blot结果显示:与对照组相比,Acrp30组自噬水平(LC3B-Ⅱ/LC3B-Ⅰ比值)24、48 h显著提高(P<0.05,P<0.01),但随时间延长,比值逐渐下降,作用72 h后LC3B-Ⅱ/LC3B-Ⅰ比值有所升高但差异无统计学意义;4流式细胞仪检测凋亡率结果显示:与对照组相比,Acrp30组和干预组48、72 h总凋亡率明显增加(P<0.05,P<0.01),与Acrp30组相比干预组72 h凋亡率显著升高(P<0.05)。结论Acrp30可抑制MCF-7细胞的增殖和迁移,诱导细胞的自噬和凋亡;抑制细胞自噬可促进Acrp30对MCF-7细胞凋亡的诱导。
Objective To explore the effect of adiponectin （Acrp30） on the proliferation, migration, autophagy, apoptosis of MCF-7 cells and investigate the relation of autophagy and apoptosis induced by adiponectin. Methods Divided the MCF-7 cells into control group （no Acrp30 added）, drug added group, then cultured for some hours . The cells viability was detected by methyl thiazolyl tetrazolium （ MTT ） assay . An appropriate concentration Acrp30 （100 ng/ml） was selected to add into the cells after culturing for some hours, the morphological changes, migration and autophagy level were quantified by invert microscope, wound-healing assay and Western blot respec- tively. The total apoptotic rate of added Acrp30 and pretreated with an autophagy inhibitor 3-methyladenine aggra- vated （MA） groups were analyzed by flow cytometry. Results @ Compared with the control group, the viability of 50,100 ng/ml cultured 72 h and 200 ng/ml cultured 48,72 h groups were lower（P 〈0. 05） ;（~） Compared with the control group,the migration lengths of Acrp30 added group 48,72 h were decreased （P 〈 0. 01 ） ;（3） The level of autophagy （autophagy associated protein LC3B-II/LC3B-I ） was up-regulated after Acrp30 added group cultured 24,48 h compared with the control group （ P 〈 0. 05, P 〈 0. 01 ）, but the ratio was reduced along with time, after 72 h, it had no difference with the control;（4） The apoptotic rate of MCF-7 cells increased after 48 h,72 h treated with 100 ng/ml Acrp30 and pretreated with MA groups （P 〈0. 05 ,P 〈0. 01 ） compared with the control group. Further- more, the 72 h apoptotic rate of （Acrp30 ＋ MA） group was increased compared with Acrp30 group （P 〈 0. 05 ）. Conclusion Acrp30 can inhibit the growth and migration of MCF-7 ceils and induce the autophagy and apoptosis. Inhibit autophagy can promote the apoptosis which induced by adiponectin in MCF-7 ceils.
出处
《安徽医科大学学报》
CAS
北大核心
2015年第9期1223-1228,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省科技攻关项目(编号:1206c0805034)
中华医学会临床医学科研专项基金项目(编号:13040420427)
关键词
脂联素
MCF-7细胞
增殖
自噬
凋亡
adiponectin
MCF-7 cell
proliferation
autophagy
apoptosis
