Study On The Mechanisms Of Calcium And HIF In Hypoxia Induced Breast Cancer Cell Line Invasion And Metastasis

Objective:1. Putting cobalt chloride into culture medium to modeling chemistry hypoxia to investigate the expression of HIF-1α(Hypoxia-inducible factor-1 alpha )in breast cancer cell under hypoxia.2. 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 breast carcinoma cell line cultured (MCF-7 and MDA-MB-231) under hypoxia.Methods:1. The effects of HIF-1αgene on the proliferation of MDA-MB-231 cell were estimated by MTT Semi quantitative reverse transcription-poly merase chain reaction(RT-PCR) is performed to detect the expression of HIF-1αmRNA in human breast cancer cell MDA-MB-231exposed during the phase of hypoxia induced by CoC12,the relations of the quantity- efficiency and the time-efficiency were analyzed.All that in order to elect the best time and concentrate to CoC12 modeling chemistry hypoxia. 2. Human breast carcinoma cell MCF-7 and MDA-MB-231 hypoxia induced by CoC12. Tests were made of control groups, CoC12groups, VPL+CoC12 groups. Semiquantitative RT-PCR was performed to detect the expression of HIF-1αmRNA in human breast carcinoma cell MCF-7 and MDA-MB-231. The protein expression level of HIF-1αwas measured by Western blotting and Immunocytochemistry. The effects of HIF-1αon the invasion and metastasis abilities of human breast cancer line MCF-7and MDA-MB-231 in vitro were explored by millicell chamber.Results:1. The best model of breast carcinoma cell hypoxia is induced by 150μmol/L CoC12 when it is 24 hours. 2. There were expressions of HIF-1αin human breast carcinoma cell MCF-7 and MDA-MB-231, But the mRNA expression level of HIF-1αand the protein expression level of HIF-1αunder hypoxia induced by CoC12 were higher than that of untreated by CoC12.The levels of mRNA and protein of HIF-1αwere suppressed significantly after treated with VPL(P<0.01). The invasive and metastatic potential in human breast cancer cells MCF-7 and MDA-MB-231 were suppressed after treated with VPL+ CoC12 than treated with CoC12 (P<0.01).,MDA-MB-231 was more significant.Conclusion:1. HIF-1αcan over-express in breast carcinoma cell inducing by CoC12.2. locking the calcium signal pathway may inhibit breast cancer celLs MCF-7 and MDA-MB-231 invasion and metastasis potential by inducing the down-regulation HIF-1αexpression.