钙信号对乳腺癌细胞HIF-1α表达及侵袭转移能力的影响.docVIP

钙信号对乳腺癌细胞HIF-1α表达及侵袭转移能力的影响 廖晓敏1，汤为学2，赖洁娟1，蒲淑萍1* （重庆医科大学 1、细胞生物及遗传教研室；2、病理生理学教研室,重庆,400016） 摘要 目的： 探讨低氧条件下钙信号对乳腺癌细胞HIF-lα表达及其侵袭转移能力的影响。方法：在培养基中加入150μmol／L CoC12模拟化学缺氧；将实验分成对照组、加CoC12组、加CoC12和钙通道阻滞剂（verapamil ，VPL）组，用逆转录-聚合酶链反应(RT-PCR)检测各组中HIF-1α的mRNA水平；蛋白印迹实验（Western blotting）和免疫细胞化学（Immunocytochemistry）检测乳腺癌细胞MCF-7和MDA-MB-231中HIF-1α蛋白的表达；Millicell小室人工重组基底膜侵袭迁移实验检测各实验组中细胞侵袭迁移能力的改变。结果：两株细胞各实验组都存在HIF-1α的mRNA转录和蛋白的表达，加CoC12组HIF-1αmRNA水平和蛋白水平较对照组显著增高（P0.01），加VPL干预后，HIF-1αmRNA水平和蛋白水平都大大降低（P0.01），MCF-7和MDA-MB-231间有显著差异（P0.01）；两株细胞的侵袭迁移能力VPL+ CoC12组较CoC12组明显降低（P0.05）,MDA-MB-231更明显（P0.01）。结论：阻断钙信号转导途径可以使HIF-1α的表达下调，进而抑制乳腺癌细胞的侵袭转移能力。 关键词： 钙信号；乳腺癌细胞；HIF-1α；侵袭转移 中图分类号：R73-37；R737.9 文献标识码：A The Effect of Calcium Signal on Expression of HIF-lα and Invasion and Metastasis of Breast Cancer Cell Line Liao Xiao-ming1, Tang Wei-xue2 ，Lai Jie Juan1，Pu Shu-ping1* （1.Department of cell biology and genetics；2. Department of Pathophysiology,Chongqing University of Medical Sciences,Chongqing 400016,China） Abstract：Objective To investigate the impact of the calcium signal on the expression of HIF-1α（Hypoxia-inducible factor-1 alpha ）and invasion and Metastasis in human 收稿日期： 作者简介：廖晓敏（1975-）,女,重庆人,硕士研究生,从事肿瘤细胞生物学研究,Tel： E-mail:liaoxm2003@163.com *通讯作者(corresponding author)：蒲淑萍 E-mail:pushuping@ breast carcinoma cell line cultured (MCF-7 and MDA-MB-231) under hypoxia . Methods Human breast carcinoma cell MCF-7 and MDA-MB-231 hypoxia induced by 150μmol/L CoC12；Semiquantitative reverse transcription-polymerase chain reaction(RT-PCR) was performed to detect the expression of HIF-1α mRNA in human breast carcinoma cell MCF-7 and MDA-MB-231；The protein expression level of HIF-1α was measured by Western blotting and Immunocytochemistry； The effects of HIF-1α on the invasion and metastasis abilities of human breast cancer line MCF-7and MDA-MB-231 in vitro were explored by millicell chamber. Results There were expressions of HIF-1α in human breast carcinoma cell MCF-7 and