钙信号对乳腺癌细胞HIF-1α表达及侵袭转移能力的影响
Effect of calcium signal on expression of HIF-1α in breast cancer cell line and invasion and metastasis of cells
廖晓敏 1汤为学 2赖洁娟 1蒲淑萍 1孙双凌1
作者信息
- 1. 重庆医科大学细胞生物学及遗传学教研室,重庆,400016
- 2. 重庆医科大学病理生理学教研室,重庆,400016
- 折叠
摘要
目的:探讨低氧条件下钙信号对乳腺癌细胞HIF-1α表达及其侵袭转移能力的影响.方法:在培养基中加入150 μmol/L CoCl2模拟化学缺氧;将实验分成对照组、CoCl2组、CoCl2和钙通道阻滞剂维拉帕米(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;蛋白印迹实验和免疫细胞化学检测乳腺癌细胞MCF-7和MDA-MB-31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,CoCl2组HIF-1αmRNA水平和蛋白水平较对照组显著增高(P<0.01),加VPL干预后,HIF-1αmRNA水平和蛋白水平都大大降低(P<0.01),MCF-7和MDA-MB-231间有显著差异(P<0.01);两株细胞的侵袭迁移能力VPL+CoCl2组较CoCl2组明显降低(P<0.05),MDA-MB-31更明显(P<0.01).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.
Abstract
AIM: To investigate the impact of the calcium signal on the expression of hypoxia-inducible factor-1 alpha( HIF-1α) in human breast carcinoma cell lines (MCF-7 and MDA-MB-231) cultured under hypoxia and the invasion and metastasis of the cells. METHODS: Human breast carcinoma cells MCF-7 and MDA-MB-231 were treated with 150 μmol/L CoCl2 to simulate the hypoxia. The cells were divided into 3 groups: control, CoCl2, and CoCl2 + verapamil (VPL) groups. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of HIF-1α mRNA in the cells. The protein expression level of HIF-1α was measured by Western Blot and immunocytochemistry. The invasion and metastasis abilities of the cells in vitro were explored by millicell chamber. RESULTS: There were expressions of HIF-1α in human breast carcinoma cells MCF-7 and MDA-MB-231. The mRNA and protein expression levels of HIF-1α under hypoxia induced by CaCl2 were higher than that of control cells ( P < 0. 01). The expression levels of mRNA and protein of HIF-1α were suppressed significantly after treatment with VPL ( P < 0. 01 ), with significant difference between MCF-7 cells and MDA-MB-231 cells (P <0. 01). The invasive and metastatic potentials of human breast cancer cells MCF-7 and MDA-MB-231 were suppressed more significantly after treatment with VPL + CoCl2 than with CoCl2 (P < 0.01) , and the suppressing effects were more significant for MDA-MB-231 than for MCF-7. CONCLUSION: Blocking the calcium signal pathway may induce the down-regulation of HIF-1α expression in breast cancer cells, so as to inhibit the invasion and metastasis potential of the cells.
关键词
钙信号/乳腺肿溜/肿瘤细胞系/HIF-1a表α达/侵袭转移Key words
calcium signal/HIF-1α/breast neoplasms/cell line/tumor/invasion and metastasis引用本文复制引用
基金项目
重庆市自然科学基金(2006BB5427)
出版年
2009
NETL
NSTL
维普
万方数据