巨噬细胞移动抑制因子对结肠癌HT-29细胞增殖和血管生成因子表达的影响

Effects of macrophage migration inhibitory factor on the proliferation and expression of angiogenesis factor in human colon carcinoma cell HT-29

目的研究巨噬细胞移动抑制因子(MIF)在体外对人结肠癌细胞HT-29增殖及VEGF和IL-8表达的影响。方法分别用25、50、100μg/L人重组MIF(rhMIF)对细胞进行处理后,采用MTT法检测细胞增殖,RT-PCR检测细胞VEGF、IL-8mRNA含量,ELISA测定细胞培养上清液中VEGF和IL-8的蛋白含量。结果rhMIF作用不同时间后,能够促进HT-29细胞生长,增殖活性随浓度增加、时间延长逐渐上升(P<0.05),呈现量—效、时—效关系。细胞VEGF和IL-8mRNA表达及上清液中VEGF和IL-8蛋白含量均明显增加,存在时间和浓度依赖性(P<0.05)。结论MIF能够促进结肠癌细胞增殖,MIF参与肿瘤血管形成有可能是通过上调VEGF和IL-8的表达发挥作用。

Objectives To investigate the effects of macrophage migration inhibitory factor on the proliferation and expression of vascular endothelial growth factor （VEGF） and interleukin-8 （IL-8） in colon carcinoma cell train HT-29. Methods The cultured HT-29 cells were treated with different concentrations of recombinant human macrophage migration inhibitory factor （rhMIF）（25, 50, 100fig/L） for different time. The proliferation of HT-29 cells was detected by MTT assay. The expression of VEGF and IL-8 mRNA in HT-29 cells were examined by real-time quantitative reverse transcription-polymerase chain reaction （RT-PCR） and the levels of VEGF and IL8 protein in the supernatants were determined by enzyme-linked immunosorbent assay （ELISA）. Results RhMIF promoted the proliferation of HT-29 cells in a concentration- and time-dependent manner （P〈0.05）, and the proliferating activity was increased with the raise of concentration and prolonging of action time. The expression of VEGF and IL-8 mRNA and the levels of VEGF and IL-8 protein in supernatants were increased gradually with the raise of concentration or action time （P〈0.05）. Conclusions MIF promotes the proliferation of colon carcinoma cells, and MIF plays a crucial role in the angiogenesis of colon cancer and the roles of MIF may be realized via raising the expression of VEGF and IL-8.