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目的探讨缺氧诱导因子1α(HIF-1α)和基因系尾型同源盒基因2(CDX2)在人结肠癌细胞株SW480和LS174T不同缺氧时间的表达及其可能的作用机制。方法(1)MTT方法检测在不同CoCl2浓度(100,150,200μmol/L)下和不同时点(24,36,48 h)时对SW480和LS174T细胞活力以及增殖的影响,筛选适宜的CoCl2作用浓度。(2)在细胞化学缺氧培养相应时段,采用半定量RT-PCR技术检测HIF-1α,Snail和CDX2的mRNA的表达变化;同时采用Western blotting技术检测CDX2的蛋白表达。结果结肠癌细胞株在不同浓度CoCl2环境下随缺氧时间的延长,细胞活力明显受到抑制。在低浓度CoCl2(100μmol/L)干预下,随着缺氧时间的延长,HIF-1α和Snail mRNA表达逐渐上升,缺氧24 h时达到高峰,CDX2 mRNA及蛋白表达水平逐渐下降。结论缺氧诱导HIF-1α过表达可通过下调CDX2而加速结直肠癌的进展。
Objective To investigate the expression of hypoxia-inducible factor-1α (HIF-1α) gene and tail-type homologue 2 (CDX2) gene in human colon cancer cell lines SW480 and LS174T under different hypoxia time and its possible mechanism. Methods (1) The effects of different concentrations of CoCl2 (100,150,200μmol / L) and different time points (24,36,48 h) on the viability and proliferation of SW480 and LS174T cells were detected by MTT assay. (2) The expression of HIF-1α, Snail and CDX2 mRNA was detected by semi-quantitative RT-PCR during the corresponding period of cytochemical hypoxia culture. Meanwhile, the protein expression of CDX2 was detected by Western blotting. Results The colon cancer cell lines with different concentrations of CoCl2 environment with hypoxia time, cell viability was significantly inhibited. Under the condition of low concentration of CoCl2 (100μmol / L), the expressions of HIF-1α and Snail mRNA increased gradually with hypoxia time, peaked 24 hours after hypoxia, and the expression of CDX2 mRNA and protein decreased gradually. Conclusion Hypoxia-induced HIF-1α overexpression can accelerate the progression of colorectal cancer by down-regulating CDX2.