摘要
目的:探讨脂联素对子宫内膜癌细胞增殖的抑制作用及AMPK/ERK信号传导通路的有关机制。方法:以10μg/ml脂联素作用子宫内膜癌Ishikawa3-H-12细胞0～60min,Western blot检测脂联素作用不同时间后细胞AMPK及ERK的磷酸化程度。脂联素作用12,24h后,分别采用RT-PCR和Western blot检测Cyclin D1 mRNA和蛋白水平。脂联素作用48h,MTT法检测细胞增殖。结果:脂联素以时间依赖方式诱导子宫内膜癌细胞AMPK活化,脂联素作用5min后明显活化,并维持至30min(F=22.749,P=0.000),AMPK抑制剂复合物C可明显抑制脂联素诱导的细胞AMPK活化。脂联素以时间依赖模式抑制子宫内膜癌细胞ERK活化,作用5min ERK活化明显受抑制,并维持至少60min(F=13.802,P=0.000),复合物C明显阻断脂联素诱导的细胞ERK活性抑制。脂联素明显抑制Cyclin D1 mRNA转录和蛋白表达(P=0.003,P=0.000),复合物C明显阻断脂联素对细胞Cyclin D1 mRNA转录和蛋白表达的抑制作用(P=0.006,P=0.000)。脂联素明显抑制细胞增殖(P=0.001),复合物C明显阻断脂联素对细胞的抑制作用(P=0.002)。结论:脂联素可能通过AMPK/ERK/Cyclin D1途径抑制子宫内膜癌细胞增殖。
Objective：To explore inhibitory effect of adiponectin on endometrial cancer cell proliferation and AMPK/ERK signal transduction mechanism.Methods：After pretreatment with 10μg/ml adiponectin for 0-60min,the phosphorylation of AMPK and ERK in endometrial cancer Ishikawa3-H-12 cell were detected by Western blot.The levels of Cyclin D1 mRNA and protein induced by 12h and 24h treatment with adiponectin were detected by RT-PCR and Western blot respectively.After 48h adiponectin treatment,proliferation of endometrial carcinoma cells was observed by methyl thiazolyl tetrazolium（MTT） assay.Results：Adiponectin induced AMPK phosphorylation of endometrial cancer cell in time-dependent manner.Activation of AMPK ocuured at 5min and lasted at least 30min（F=22.749,P=0.000）.The AMPK activation induced by Adiponectin was blocked by AMPK inhibitor comfound C.Adiponectin inhibited ERK phosphorylation in time-dependent manner.ERK activation decreased at 5min and continued to 60min at least（F=13.802,P=0.000）.Comfound C abrogated ERK inhibition of adiponectin.Both Cyclin D1 mRNA and protein expressions were inhibited by adiponectin（P=0.003,P=0.000 respectively）.These efftects were blocked by comfound C（P=0.006,P=0.000 respectively）.After adiponectin treatment,proliferation of endometrial cancer cells was lower than that of control group（P=0.001）.Comfound C abrogated the inhibitory effect induced by adiponectin on the proliferation of endometrial cancer cells（P=0.002）.Conclusion：Adiponectin maybe inhibit proliferation of endometrial cancer cells through AMPK/ERK/Cyclin D1 signal pathway.
出处
《现代妇产科进展》
CSCD
2012年第11期861-864,共4页
Progress in Obstetrics and Gynecology
基金
天津市卫生局科技基金重点项目(No:2011KR04)
