脂联素对子宫内膜癌细胞增殖凋亡的影响

目的:研究脂联素对子宫内膜癌细胞增殖凋亡的影响及其信号的传导。方法:免疫细胞化学方法及RT-PCR法检测子宫内膜癌Ishikawa3-H-12细胞AdipoR1/R2蛋白及mRNA的表达,不同浓度脂联素(2.5,5,10,20μg/ml)作用子宫内膜癌细胞30min,Western blot检测AMPK磷酸化程度,不同浓度脂联素作用细胞48h及AMPK抑制剂(复合物C)预处理后脂联素再作用48h后,MTT试验及流式细胞仪检测不同处理组细胞的增殖及凋亡。结果:子宫内膜癌Ishikawa3-H-12细胞AdipoR1/R2蛋白呈棕黄色颗粒状表达,并可见AdipoR1/R2基因表达。除2.5μg/ml浓度外,其余浓度组脂联素均显著诱导AMPK磷酸化,不同浓度组AMPK活化差异有统计学意义(F=27.985,P0.01),脂联素以浓度依赖模式诱导子宫内膜癌细胞AMPK活化。脂联素以浓度依赖模式显著抑制子宫内膜癌细胞增殖(F=13.322,P0.01),诱导子宫内膜癌细胞凋亡(F=46.826,P0.01),复合物C可以阻断脂联素诱导的细胞增殖抑制及凋亡。结论:脂联素可能通过影响AMPK信号通路抑制子宫内膜癌细胞增殖,促进凋亡。

Effects of adiponectin on proliferation and apoptosis of endometrial cancer cells

Objective:To explore the effects of adiponectin on proliferation and apoptosis of endometrial cancer cell as well as its signal pathway.Methods:Adipo R1/R2 mRNA and protein were detected by RT-PCR and immunocytochemistry in endometrial cancer cell line Ishikawa3-H-12. Western blotting was performed to determine the phosphorylation level of AMPK(p-AMPK).Cell proliferation was evaluated by methyl thiazolyl tetrazolium(MTT) assay and apoptosis was assessed by flow cytometry. Pretreatment of the endometrial cancer lines with AMPK inhibitor(compound C) or blank received either different concentrations of adiponectin(2.5,5,10,20ug/ml) and were collected for 30min or 24 hours respectively.The cells were collected in 30min for phosphorylation level of AMPK assessment and in 24 hours for cell proliferation and apoptosis evaluation.Results:Expressions of adipo R1/R2 in Ishikawa3-H-12 cell line were demonstrated.Stimulation of the Ishikawa cells with adiponectin at different concentrations resulted in a dose-dependent activation of AMPK(F=27.985,P<0.01).After adiponectin treatment,the OD570 were all lower than that of the control group in a dose-dependent manner(F=13.322,P<0.01).The apoptotic rates of adiponectin groups were higher than that of the control group in a dose-dependent manner(F=46.826,P<0.01).The effects of proliferation inhibition and apoptosis enhance of adiponectin were blocked by compound C.Conclusion:Proliferation inhibition and apoptosis promotion induced by adiponectin in endometrial cancer cells may be mediated through AMPK signal pathway.