1,25二羟维生素D3对卵巢癌A2780细胞自噬和凋亡的影响  

Effects of 1,25-dihydroxyvitamin D3 on autophagy and apoptosis of ovarian cancer A2780 cells

作　　者：刘华 周玲玲 魏丹丹 徐宜艳 张慧 Liu Hua;Zhou Lingling;Wei Dandan;Xu Yiyan;Zhang Hui(Obstetrics and Gynecology,People’s Hospital of Yicheng City,Yicheng 441400,China;Obstetrics and Gynecology,Xiangyang Central Hospital,Xiangyang 441021,China)

机构地区：[1]湖北宜城市人民医院妇产科,宜城441400 [2]湖北襄阳市中心医院湖北文理学院附属医院妇产科,襄阳441021

出　　处：《广西医科大学学报》2020年第8期1449-1455,共7页Journal of Guangxi Medical University

基　　金：湖北省自然科学基金资助项目(No.2017CFB325)。

摘　　要：目的:探讨1,25二羟维生素D3(VD3)对卵巢癌A2780细胞自噬和凋亡的影响。方法:将细胞随机分为对照组、低、中、高剂量组、高剂量+IGF-1组。低、中、高剂量组卵巢癌A2780细胞分别用2.5 nmol/L、5 nmol/L、10 nmol/L VD3处理24 h,高剂量+IGF-1组用10 nmol/L VD3和10 ng/mL IGF-1处理24 h。BrdU染色检测细胞增殖;流式细胞术检测细胞凋亡;免疫荧光检测LC3的含量;蛋白印迹法检测凋亡相关蛋白(Caspase3、Caspase9)、自噬相关蛋白(ATG7、Beclin1、LC3)和磷脂酰肌醇激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白的表达水平。结果:与对照组比较,中、高剂量组BrdU阳性细胞数显著减少(P<0.05);细胞凋亡率、活化的Caspase3/Caspase3和活化的Caspase9/Caspase9蛋白表达的比值、LC3水平、ATG7和Beclin1蛋白表达水平均显著升高(P<0.05);低、中、高剂量组LC3Ⅱ/LC3Ⅰ蛋白表达水平比值显著升高(P<0.05);PI3K、AKT和mTOR磷酸化水平显著降低(P<0.05)。激活PI3K/AKT/mTOR通路逆转VD3对A2780细胞的作用(P<0.05)。结论:VD3可抑制PI3K/AKT/mTOR通路的磷酸化,诱导卵巢癌A2780细胞自噬和凋亡。Objective:To investigate the effects of 1,25-dihydroxyvitamin D3(VD3)on autophagy and apoptosis of ovarian cancer A2780 cells.Methods:The cells were randomly divided into control group,low dose group,middle dose group,high dose group and high dose+IGF-1 group.Ovarian cancer A2780 cells in low,middle and high dose groups were treated with 2.5 nmol/L,5 nmol/L and 10 nmol/L VD3 for 24 hours,while those in high dose+IGF-1 group were treated with 10 nmol/L VD3 and 10 ng/mL IGF-1 for 24 hours.Cell proliferation was detected by BrdU staining.Apoptosis was detected by flow cytometry.The content of LC3 was detected by immunofluorescence.The expression levels of apoptosis-related proteins(Caspase3,Caspase9),autophagy-related proteins(ATG7,Beclin1,LC3)and phosphatidylinositol kinase(PI3K)/protein kinase B(AKT)/mammalian rapamycin target protein(mTOR)pathway were detected by Western blotting.Results:Compared with the control group,the number of BrdU positive cells in the middle and high dose groups decreased significantly(P<0.05).The apoptosis rate,the ratio of activated Caspase3/Caspase3the ratio of activated Caspase9/Caspase9 protein,LC3 level,ATG7 and Beclin1 protein expression increased significantly(P<0.05).The ratio of LC3II/LC3I protein expression increased significantly in low,middle and high dose groups(P<0.05),and the phosphorylation levels of PI3K,AKT and mTOR decreased significantly(P<0.05).Activation of PI3K/AKT/mTOR pathway reversed the effect of VD3 on A2780 cells(P<0.05).Conclusion:VD3 can inhibit the phosphorylation of PI3K/AKT/mTOR pathway and induce autophagy and apoptosis in ovarian cancer A2780 cells.

关 键 词：1 25二羟维生素D3 PI3K/AKT/MTOR 卵巢癌 自噬 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]