低氧环境对人上皮性卵巢癌细胞株上皮间质转化及侵袭能力的影响及机制探讨  

The effect of hypoxia-inducible factor-1α on epithelial-mesenchymal transition and invasion in human epithelial ovarian cancer cells

作　　者：曹珊[1] 闫晓楠[1] 展艳萍[1] 徐春琳[1] 王惠兰[1] 

机构地区：[1]河北医科大学第二医院妇产科,石家庄050000

出　　处：《现代妇产科进展》2014年第11期864-867,共4页Progress in Obstetrics and Gynecology

摘　　要：目的:探讨低氧环境对人卵巢癌细胞上皮间质转化(EMT)和侵袭能力的影响及可能机制。方法:培养人卵巢癌细胞株SKOV-3、3AO,将每株细胞分为对照组、Co Cl2(模拟低氧环境)组、VPL(钙离子拮抗剂)组和VPL+Co Cl2组。采用四甲基偶氮唑蓝(MTT)比色法检测Co Cl2对SKOV-3、3AO细胞生长情况的影响;倒置相差显微镜下观察低氧后细胞形态的改变;采用Western blot法检测SKOV-3、3AO细胞中缺氧诱导因子-1α(HIF-1α)、转录因子Twist、上皮型钙黏蛋白(E-cad)及神经型钙黏蛋白(N-cad)表达的变化;体外侵袭实验穿膜小室(Transwell小室)检测细胞的侵袭能力。结果:Co Cl2作用后,两种细胞的存活率均较对照组明显降低(P<0.05)。低氧环境下,部分细胞伸出伪足,呈多角形,细胞排列松散、紊乱。Co Cl2能体外模拟细胞低氧环境,诱导HIF-1α蛋白高表达。SKOV-3、3AO细胞中,Co C12组中HIF-1α、Twist、N-cad蛋白表达明显高于对照组、Co C12+VPL组(P<0.01,P<0.05);E-cad蛋白表达明显低于对照组、Co C12+VPL组(P<0.01,P<0.05);VPL组与对照组相比,各蛋白表达的变化无明显差异(P>0.05)。Co C12组两种细胞的穿膜数均显著多于对照组、Co C12+VPL组(P<0.01,P<0.05)。结论:钙信号途径参与了低氧环境下HIF-1α调节卵巢癌SKOV-3、3AO细胞中Twist的表达,从而诱导这两种细胞EMT的发生并增强其侵袭能力。Objective：To inventigate the effect of hypoxia on regulating epithelial mes-enchymal transition （EMT） and invasion in human ovarian cancer cells and its possible mecha-nism. Methods：The human ovarian cancer cell lines of SKOV-3 and 3AO were cultured and as-signed to control group,CoCl2 （Simulating the hypoxic environment） group,VPL（calcium an-tagonists）group and CoCl2＋VPL group,respectively. The effect of CoCl2 on cell growth was de-tectd by methyl thiazolyl tetrazolium （MTT） assay. The morphological changes of SKOV-3 and 3AO were observed by the inverted phase contrast microscope. The expression of HIF-1α pro-tein by different concentration of CoCl2 in various times,as well as the expression of the tran-scription factors Twist,E-cad and N-cad protein,were detected by Western blotting. The cell in-vasion was tested by Transwell invasion assay. Results：The survival rates of SKOV-3 and 3AO cells in CoCl2 group were significantly lower than that in the control group （P〈0. 05）. In hy-poxia environment,SKOV-3 and 3AO cells lost polarity and partially presented the characteris-tic of polygons cells with pseudopods. Cell-cell junctions were receded and the cells were dis-persed. The hypoxic environment in vitro could be created by CoCl2 ,which could induce higher expression of HIF-1α protein. Compared with the control group,higher expression of HIF-1α, Twist and N-cadherin protein were observed in CoCl2 group （P〈0. 05）. VPL reversed the effect of CoCl2 on increasing the expression of HIF-1α,Twist and N-cadherin,there was significant difference comparing CoCl2 group with CoCl2＋VPL group. Whereas the expression of E-cadherin was reduced in CoCl2 group,compared with the control group（ P 〈0. 01）. VPL reversed the effect of CoCl2 on decreasing the expression of E-cadherin,there was significant difference com-paring CoCl2 group with CoCl2 ＋VPL group. But there was no significant difference in the ex-pression of HIF-1α, Twist, E-cadherin and N-cadherin between VPL group and th

关 键 词：上皮性卵巢癌 低氧诱导因子-1Α 钙信号 上皮间质转化 侵袭转移 

分 类 号：R737.33[医药卫生—肿瘤]