磷脂酰肌醇-3-激酶/蛋白激酶B和丝裂原活化蛋白激酶/细胞信号调节激酶1/2信号通路抑制剂对乳腺癌细胞缺氧诱导因子-2α表达的影响  

Effects of phosphatidylinositol-3-kinase/protein kinase B and mitogen activated protein kinase/cell signal regulated kinase 1/2 signaling pathways inhibitors on the expression of hypoxia inducible factor-2αin breast cancer cells

作　　者：李永真[1] 王志慧 李娜[1] 宋颖[1] 韩正华 千新来[1] 原志庆[1] 陆漫漫 李思琦 LI Yongzhen;WANG Zhihui;LI Na;SONG Ying;HAN Zhenghua;QIAN Xinlai;YUAN Zhiqing;LU Manman;LI Siqi(Department of Pathology,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Pathdogy,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;College of Sanquan,Xinxiang Medical Univer sity,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院病理学系,河南新乡453003 [2]新乡医学院第三附属医院,河南新乡453003 [3]新乡医学院三全学院,河南新乡453003

出　　处：《新乡医学院学报》2021年第4期308-312,共5页Journal of Xinxiang Medical University

基　　金：河南省科技厅科技攻关资助项目(编号:202102310424);河南省教育厅高等学校重点科研项目(编号:21A310014);新乡医学院基础医学院科研培育项目(编号:JCYXYKY202017)。

摘　　要：目的探讨磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)和丝裂原活化蛋白激酶/细胞信号调节激酶1/2(MEK/ERK1/2)信号通路抑制剂对缺氧诱导的乳腺癌MCF-7细胞中缺氧诱导因子-2α(HIF-2α)表达的影响。方法乳腺癌MCF-7细胞常规培养24 h后,分为常氧组、缺氧组、缺氧+低剂量LY294002组、缺氧+高剂量LY294002组,缺氧+低剂量U0126组和缺氧+高剂量U0126组。常氧组细胞使用含生理盐水的RPMI 1640培养基,缺氧组细胞使用含100μmol·L-1氯化钴的RPMI 1640培养基,缺氧+低剂量LY294002组细胞使用含100μmol·L-1氯化钴和4μmol·L-1 LY294002的RPMI 1640培养基,缺氧+高剂量LY294002组细胞使用含100μmol·L-1氯化钴和40μmol·L-1 LY294002的RPMI 1640培养基,缺氧+低剂量U0126组细胞使用含100μmol·L-1氯化钴和2μmol·L-1 U0126的RPMI 1640培养基,缺氧+高剂量U0126组细胞应用含8μmol·L-1 U0126的RPMI 1640培养基,继续培养3 h。采用实时荧光定量聚合酶链式反应检测各组细胞HIF-2αmRNA的表达;采用Western blot法检测各组细胞HIF-2α蛋白、磷酸化Akt(p-Akt)和磷酸化ERK1/2(p-ERK1/2)蛋白的表达。结果缺氧组细胞中HIF-2αmRNA相对表达量及HIF-2α蛋白、p-Akt蛋白和p-ERK1/2蛋白相对表达量高于常氧组(P<0.05);缺氧+低剂量LY294002组、缺氧+高剂量LY294002组、缺氧+低剂量U0126组、缺氧+高剂量U0126组细胞中HIF-2αmRNA相对表达量及HIF-2α蛋白、p-Akt蛋白和p-ERK1/2蛋白相对表达量低于缺氧组(P<0.05)。缺氧+高剂量LY294002组细胞中HIF-2αmRNA相对表达量及HIF-2α蛋白、p-Akt蛋白和p-ERK1/2蛋白相对表达量低于缺氧+低剂量LY294002组(P<0.05);缺氧+高剂量U0126组细胞中HIF-2αmRNA相对表达量及HIF-2α蛋白、p-Akt蛋白和p-ERK1/2蛋白相对表达量低于缺氧+低剂量U0126组(P<0.05)。结论PI3K/Akt和MEK/ERK1/2信号通路可能是缺氧诱导HIF-2α表达的上游信号通路,在缺氧条件下对HIF-2α具有正向调节作用,在乳腺癌的发生、�Objective To investigate the effects of phosphatidylinositol-3-kinase/protein kinase B(PI3K/Akt)and mitogen activated protein kinase/cell signal regulated kinase 1/2(MEK/ERK1/2)signaling pathways inhibitors on the expre-ssion of hypoxia inducible factor-2α(HIF-2α)in hypoxia-induced breast cancer MCF-7 cells.Methods The breast cancer MCF-7 cells after routinely cultured for 24 h were divided into normoxia group,hypoxia group,hypoxia+low-dose LY294002 group,hypoxia+high-dose LY294002 group,hypoxia+low-dose U0126 group and hypoxia+high-dose U0126 group.The cells in the normoxia group were cultured with RPMI 1640 medium containing normal saline,the cells in the hypoxia group were cultured with RPMI 1640 medium containing 100μmol·L-1 CoCl 2,the cells in the hypoxia+low-dose LY294002 group were cultured with RPMI 1640 medium containing 100μmol·L-1 CoCl 2 and 4μmol·L-1 LY294002,the cells in the hypoxia+high-dose LY294002 group were cultured with RPMI 1640 medium containing 100μmol·L-1 CoCl 2 and 40μmol·L-1 LY294002,the cells in the hypoxia+low-dose U0126 group were cultured with RPMI 1640 medium containing 100μmol·L-1 CoCl 2 and 2μmol·L-1 U0126,and the cells in the hypoxia+high-dose U0126 group were cultured with RPMI 1640 medium containing 100μmol·L-1 CoCl 2 and 8μmol·L-1 U0126,the culture was continued for 3 h.The expression of HIF-2αmRNA was detected by real-time quantitative polymerase chain reaction,the expression of HIF-2αprotein,phosphorylated Akt(p-Akt)protein and phosphorylated ERK1/2(p-ERK1/2)protein were detected by Western blot.Results The relative expression levels of HIF-2αmRNA,HIF-2αprotein,p-Akt protein and p-ERK1/2 protein in the hypo xia group were higher than those in the normoxia group(P<0.05);the relative expression levels of HIF-2αmRNA,HIF-2αprotein,p-Akt protein and p-ERK1/2 protein in the hypoxia+low-dose LY294002 group,hypoxia+high-dose LY294002 group,hypoxia+low-dose U0126 group and hypoxia+high-dose U0126 group were lower than those in the hypoxia group(P<0.05).The re

关 键 词：乳腺癌 缺氧诱导因子-2Α 磷脂酰肌醇-3-激酶/蛋白激酶B通路 丝裂原活化蛋白激酶/细胞信号调节激酶1/2信号通路 

分 类 号：R737.9[医药卫生—肿瘤]