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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,P<0.01),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,P<0.01),诱导子宫内膜癌细胞凋亡(F=46.826,P<0.01),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。
Objective: To study the effect of adiponectin on proliferation and apoptosis of endometrial carcinoma cells and the signal transduction. Methods: The expressions of AdipoR1 / R2 protein and mRNA in Ishikawa 3-H-12 cells were detected by immunocytochemistry and RT-PCR. Adiponectin (2.5, 5, 10, 20μg / ml) The phosphorylation of AMPK was detected by Western blot and Western blot analysis. The adiponectin was pretreated with different concentration of adiponectin for 48 h and pretreatment with AMPK inhibitor (compound C) for 48 h. MTT assay and flow cytometry Treatment group cells proliferation and apoptosis. Results: The AdipoR1 / R2 protein of Ishikawa3-H-12 cells was expressed as brown granules and the expression of AdipoR1 / R2 gene was observed. In addition to 2.5μg / ml concentration, the other concentrations of adiponectin significantly induced AMPK phosphorylation, AMPK activation in different concentrations were statistically significant difference (F = 27.985, P <0.01), adiponectin concentration-dependent manner to induce uterine Endometrial cancer cells activate AMPK. Adiponectin significantly inhibited the proliferation of endometrial cancer cells in a dose-dependent manner (F = 13.322, P <0.01), and induced the apoptosis of endometrial cancer cells (F = 46.826, P <0.01) Tetrandrine induced cell proliferation inhibition and apoptosis. Conclusion: Adiponectin may inhibit apoptosis of endometrial cancer cells through affecting AMPK signaling pathway.