缺氧环境对宫颈腺癌细胞增殖和迁移的影响及其作用机制  

Effects of hypoxia on proliferation and migration of cervical adenocarcinoma cells and its mechanism

作　　者：刘艳 李月洁 黄小环[2] 龚莉 周铁军 LIU Yan;LI Yuejie;HUANG Xiaohuan;GONG Li;ZHOU Tiejun(The Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院,四川泸州646000 [2]重庆三峡医药高等专科学校

出　　处：《山东医药》2019年第31期46-50,共5页Shandong Medical Journal

基　　金：四川省科学技术与泸州市人民政府、泸州医学院联合科研专项资金计划项目(14JC01773-LH72);泸州医学院附属医院科研课题计划[(2014)71-180]

摘　　要：目的探讨缺氧环境对宫颈腺癌细胞增殖和迁移的影响及其作用机制。方法将传3代、对数生长期、生长状态良好的宫颈腺癌HeLa细胞接种于96孔板,分别给予50、100、150、200、250μmol/L氯化钴(CoCl 2)干预,另设对照孔,不予CoCl 2干预。干预24、48、72 h,收集细胞,采用MTT法检测各孔细胞增殖抑制率。选取细胞增殖抑制率最低的CoCl 2浓度作为后续构建细胞缺氧模型的最佳浓度。将HeLa细胞接种于6孔板,随机分为观察组和对照组,观察组通过CoCl 2构建细胞缺氧模型,对照组常规培养,显微镜下观察继续培养0、48 h划痕距离,计算细胞迁移率。将HeLa细胞接种于6孔板,通过CoCl 2构建细胞缺氧模型,分别于CoCl 2干预0、24、48、72 h收集细胞,采用RT-PCR、Western blotting法检测缺氧诱导因子2α(HIF-2α)、血管内皮钙黏蛋白(VE-cadherin)mRNA和蛋白表达。结果50、100μmol/L CoCl 2干预24 h能够促进HeLa细胞增殖(P均<0.05),且以100μmol/L时效果更强,故后续选择100μmol/L CoCl 2构建细胞缺氧模型。150、200、250μmol/L CoCl 2干预24 h即可表现出对HeLa细胞增殖的抑制作用,且随着干预时间延长,细胞增殖抑制作用越来越明显(P均<0.05)。观察组与对照组细胞迁移率分别为(61.4±4.8)%、(37.9±3.6)%,两组比较P<0.01。缺氧环境下,HeLa细胞HIF-2α、VE-cadherin mRNA和蛋白相对表达量均明显升高(P均<0.05),但后者表达随着缺氧时间延长有所降低。Pearson直线相关分析显示,缺氧环境下HeLa细胞HIF-2α表达与VE-cadherin表达呈正相关关系(P均<0.05)。结论一定程度缺氧能够促进宫颈腺癌细胞增殖和迁移能力,其机制可能与缺氧环境能够增强HIF-2α、VE-cadherin表达有关。Objective To investigate the effects of hypoxia environment on the proliferation and migration of cervical adenocarcinoma cells and its mechanism.Methods The cervical adenocarcinoma HeLa cells of the third generation,in logarithmic growth phase and good growth state were inoculated into 96-well plates and treated with 50,100,150,200,and 250μmol/L CoCl 2.Meanwhile,the control group was set and not treated with CoCl 2.The cells were harvested at 24,48 and 72 h,and the proliferation inhibition rate of each well was detected by MTT.The concentration of CoCl 2 with the lowest cell proliferation inhibition rate was selected as the optimal concentration for the subsequent construction of cell hypoxia model.HeLa cells were inoculated into 6-well plates and randomly divided into the observation group and control group;in the observation group,the hypoxia models were constructed with CoCl 2,and the cells in the control group were routinely cultured.The scratch distance was observed under microscope at 0 and 48 h,and the cell migration rate was calculated.HeLa cells were inoculated into 6-well plates,and the hypoxia model was constructed by CoCl 2.The cells were harvested at 0,24,48,and 72 h after CoCl 2 intervention.RT-PCR and Western blotting were used to detect the hypoxia-inducible factor-2α(HIF-2α)and vascular endothelium-cadherin(VE-cadherin)expression of gene and protein.Results Fifty and 100μmol/L CoCl 2 could promote the proliferation of HeLa cells(both P<0.05),and the effect was stronger at 100μmol/L.Therefore,the cell hypoxia model was constructed by selecting 100μmol/L CoCl 2.The study also found that 150,200,and 250μmol/L CoCl 2 intervention for 24 h showed the inhibition of HeLa cell proliferation,and the cell proliferation inhibition effect was more significant over intervention time(P<0.05).The cell migration rates of the observation group and the control group were(61.4±4.8)%and(37.9±3.6)%(both P<0.01).The relative mRNA and protein expression of HIF-2αand VE-cadherin in HeLa cells increased under

关 键 词：宫颈腺癌 缺氧环境 缺氧诱导因子2α 血管内皮钙黏蛋白 细胞增殖 细胞迁移 

分 类 号：R737.33[医药卫生—肿瘤]