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HIF-2α/Notch3通路介导CoCl2诱导的乳腺癌MCF-7细胞迁移和侵袭

王建国, 袁磊^*

漯河医学高等专科学校分子医学实验室，漯河 462002

摘要

本研究旨在探究HIF-2α和Notch3在CoCl2诱导的乳腺癌MCF-7细胞迁移和侵袭中的作用。使用CoCl2于体外建立化学性低氧模型；采用shRNA技术沉默MCF-7细胞中HIF-2α和Notch3基因；采用RT-PCR法检测MCF-7细胞中HIF-2α、Notch3和Hey1的mRNA水平；采用Western blot方法检测MCF-7细胞中HIF-2α、Notch3、Hey1、Snail和E-cadherin蛋白的表达水平；采用划痕实验和Transwell实验分别检测CoCl2诱导的MCF-7细胞迁移和侵袭。结果显示，CoCl2处理可使MCF细胞中HIF-2α、Notch3、Hey1和Snail蛋白表达水平升高(P < 0.05)，E-cadherin蛋白表达水平下降(P < 0.05)，促进MCF-7细胞迁移和侵袭(P < 0.05)；沉默HIF-2α基因表达可抑制CoCl2诱导的Notch3和Hey1的mRNA和蛋白表达(P < 0.05)；抑制Notch3基因表达后，CoCl2诱导的MCF-7细胞迁移和侵袭以及CoCl2对Snail和E-cadherin蛋白表达的调节随之受到抑制(P < 0.05)。以上结果表明，在CoCl2化学性低氧环境下，HIF-2α可通过强化Notch3信号通路，进而导致Snail蛋白水平升高和E-cadherin蛋白水平下降，最终提高乳腺癌MCF-7细胞的迁移和侵袭能力。

关键词： 低氧诱导因子-2α; Notch3; 细胞迁移; MCF-7细胞; 侵袭

分类号：R737.9

[HIF-2α/Notch3 pathway mediates CoCl2-induced migration and invasion in human breast cancer MCF-7 cells.] [Article in Chinese]

WANG Jian-Guo, YUAN Lei^*

Laboratory of Molecular Biology, Luohe Medical College, Luohe 462002, China

Abstract

The aim of this study is to investigate the effects of hypoxia inducible factor-2α (HIF-2α) and Notch3 on CoCl2-induced migration and invasion of human breast cancer cell line MCF-7. MCF-7 cells were exposed to normoxia (21% O2) or chemical hypoxia (21% O2 plus CoCl2). Short hairpin RNA (shRNA) was used to knock down HIF-2α and Notch3 in MCF-7 cells. The mRNA expression levels of HIF-2α, Notch3 and Hey1 were measured by RT-PCR. Western blot was performed to determine the protein expression levels of HIF-2α, Notch3, Hey1, Snail and E-cadherin. CoCl2 treatment resulted in higher protein expression levels of HIF-2α, Notch3, Hey1, Snail (P < 0.05) and lower levels of E-cadherin (P < 0.05), and promoted migration and invasion of MCF-7 cells (P < 0.05). shRNA-HIF-2α suppressed CoCl2-induced mRNA expression of Notch3 and Hey1. Notch3 knockdown down-regulated Snail and up-regulated E-cadherin at protein level under simulated hypoxia (P < 0.05), and inhibited CoCl2-induced migration and invasion of MCF-7 cells (P < 0.05). In conclusion, our data provide evidence that HIF-2α may promote the migration and invasion of MCF-7 cells under chemical hypoxic conditions by potentiating Notch3 pathway.

Key words: hypoxia inducible factor-2α; Notch3; cell migration; MCF-7 cells; invasion

收稿日期：2016-03-16　　录用日期：2016-05-03

通讯作者：袁磊　　E-mail: yl_fzyx@163.com

引用本文：

王建国, 袁磊. HIF-2α/Notch3通路介导CoCl2诱导的乳腺癌MCF-7细胞迁移和侵袭[J]. 生理学报 2016; 68 (6): 783-789.

WANG Jian-Guo, YUAN Lei. [HIF-2α/Notch3 pathway mediates CoCl2-induced migration and invasion in human breast cancer MCF-7 cells.] [Article in Chinese]. Acta Physiol Sin 2016; 68 (6): 783-789 (in Chinese with English abstract).