青蒿琥酯对子宫内膜癌细胞增殖抑制的影响

Effect of artesunate on proliferation inhibition of endometrial carcinoma cells

目的研究青蒿琥酯对子宫内膜癌细胞增殖抑制及腺苷酸活化蛋白激酶(AMPK)/哺乳类雷帕霉素靶蛋白(mTOR)/核糖体蛋白S6激酶1(S6K1)信号通路的影响。方法体外培养子宫内膜癌Ishikawa细胞,低、中、高剂量实验组分别以60,90,120μg·mL^-1青蒿琥酯处理,对照组用等量生理盐水处理。各组分别干预24,48,72 h。以倒置显微镜观察细胞形态;用活细胞计数(CCK-8)法及平板克隆形成实验检测细胞增殖情况;用蛋白质印迹(Wb)法检测磷酸化AMPK(p-AMPK)、磷酸化mTOR(p-mTOR)、磷酸化S6K1(p-S6K1)蛋白表达水平。结果干预72 h时,低、中、高剂量实验组子宫内膜癌Ishikawa细胞增殖抑制率分别为(41.37±2.86)%,(53.24±4.13)%和(76.63±6.21)%;干预72 h时,对照组和低、中、高剂量实验组子宫内膜癌Ishikawa细胞克隆形成率分别为(72.12±4.34)%,(46.51±3.63)%,(31.02±2.94)%,(14.56±4.22)%。随干预时间延长和青蒿琥酯浓度增大,低、中、高剂量实验组子宫内膜癌Ishikawa细胞增殖抑制率逐渐升高(P<0.05);与对照组相比,低、中、高剂量实验组子宫内膜癌Ishikawa细胞p-AMPK蛋白相对表达量显著升高,而克隆形成率与p-mTOR、p-S6K1蛋白相对表达量降低,且呈浓度依赖性(均P<0.05)。结论青蒿琥酯可激活AMPK,通过AMPK/mTOR/S6K1信号通路抑制子宫内膜癌Ishikawa细胞增殖,发挥抗肿瘤作用。

Objective To investigate the effect of artesunate on proliferation inhibition of endometrial carcinoma cells and adenylate-activated protein kinase(AMPK)/mammalian rapamycin target protein(mTOR)/ribosomal protein S6 kinase 1(S6K1)signaling pathway.Methods The Ishikawa cells of endometrial carcinoma were cultured in vitro,and the Exp-L,Exp-M,Exp-H groups were treated with 60,90,120μg·mL^-1 artesunate,control group was treated with the same amount of normal saline,each group was treated for 24,48 and 72 h.The morphology of the cells were observed with an inverted microscope;the proliferation of cells was detected by cell counting kit(CCK-8)method and plate colony formation assay;Western blotting(Wb)method was used to detect the protein expression levels of phosphorylated AMPK(p-AMPK),phosphorylated mTOR(p-mTOR)and phosphorylated S6K1(p-S6K1).Results At 72 h after intervention,the proliferation inhibition rates of endometrial carcinoma Ishikawa cells in Exp-L,Exp-M,Exp-H groups were(41.37±2.86)%,(53.24±4.13)and(76.63±6.21)%.At 72 h after intervention,the formation rates of endometrial carcinoma Ishikawa cells in control group and Exp-L,Exp-M,Exp-H groups were(72.12±4.34)%,(46.51±3.63)%,(31.02±2.94)%,(14.56±4.22)%.With the prolongation of intervention time and the increase of artesunate concentration,the proliferation inhibition rate of endometrial carcinoma Ishikawa cells of Exp-L,Exp-M,Exp-H groups increased gradually(P<0.05).Compared with control group,the relative expression of p-AMPK protein of Exp-L,Exp-M,Exp-H groups were significantly increased,the clone formation rate and the relative expression of p-mTOR and p-S6K1 protein were significantly decreased in a concentration-dependent manner(all P<0.05).Conclusion Artesunate can activate AMPK and inhibit the proliferation of endometrial carcinoma ishikawa cells through AMPK/mTOR/S6K1 signaling pathway,and play an anti-tumor effect.