ISO-1对大肠癌细胞侵袭的影响  

The effects of ISO-1 on the invasion of colorectal cancer cell line

作　　者：陈萌[1] 何兴祥[2] 刘成勇[1] 苏杭[3] 郭海波[3] 

机构地区：[1]广州医学院第二附属医院消化科,广州510260 [2]广东药学院临床医学院内科教研室,广州510330 [3]广州医学院第二附属医院临床分子医学实验中心,广州510260

出　　处：《广东医学》2008年第5期734-737,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30470435);广东省自然科学基金资助项目(编号:06022450;7101731);广东药学院科研启动基金资助项目(编号:2006LCY07);广州市科学技术局科技攻关计划项目(编号:2006Z3-E0201)

摘　　要：目的研究选择性MIF互变异构酶活性抑制剂ISO-1对人大肠癌细胞侵袭的影响并探讨其可能的机制。方法实验组用0．01-100μmol／L的ISO-1作用于大肠癌细胞Lovo，对照组为相应浓度的DMSO处理；MIF互变异构酶活性通过L-多巴色素甲酯测定；微孔迁移法检测ISO-1对Lovo细胞体外侵袭的影响；ELISA法测定ISO-1作用后培养上清中MIF蛋白水平；逆转录聚合酶链反应（RT-PCR）检测ISO-1对Lovo细胞MMP-9和IL-8 mRNA表达的影响。结果较高浓度ISO-1（10-100μmol／L）均能明显抑制MIF互变异构酶活性，但不影响细胞外分泌MIF蛋白水平；100μmol／L ISO-1作用24h Lovo细胞穿透聚碳酸酯膜显著减少，与对照组比较差异有显著性（153±13vs204±10，P〈0．05）；100μmol／L ISO-1作用24h Lovo细胞表达MMP-9和IL-8 mRNA水平显著降低，与对照组比较差异有显著性（MMP-9 mRNA：0．5187±0．072vs0．6757±0．026．P〈0．05；IL-8 mRNA：0．1541±0．019vs0．2081±0．030，P〈0．05）。结论选择性MIF活性抑制剂ISO-1降低了Lovo细胞的侵袭，其可能机制为ISO-1抑制了MIF互变异构酶活性并下调MMP-9和IL-8的表达。Objective To investigate the effects of ISO - 1, a selective MIF tautomerase activity inhibitor, on the invasion of human colorectal cancer cell llne and its probable mechanisms. Methods The colorectal cancer cell line Lovo cells were treated with serial concentrations of ISO - 1 （0.01 mol/L to 100 μmol/L） ,while those in the control group were treated with the corresponding concentration of dimethylsulfoxide （DMSO）. The tautomerase activity of MIF was measured using L - dopachrome methyl ester. Microporous migration assay was performed to determine the effect of ISO - 1 on the invasion of Lovo cells. ELISA was used to determined MIF protein levels. The expression of MMP - 9 （ matrix metalloproteinase - 9） and IL - 8 （ Interleukin - 8 ） mRNA was detected by reverse transcription polymerase chain reaction （ RT - PCR）. Results Higher concentrations of ISO - 1 （ 10mol/L to 100μmol/L） significantly inhibited tautomerase activities of MIF ,while extracellular secretory MIF protein levels were not significantly reduced. In comparison with the control, 100μmol/L ISO - 1 significantly reduced the average number of cells penetrating polycarbonates. The results were significantly different （ 153±13 vs 204±10, P 〈 0. 05 ）. The expression of MMP - 9 and IL - 8 mRNA was significantly reduced after 100 μmol/L ISO - 1 treatment for 24 hours. The results were significantly different （MMP -9 mRNA： 0. 518 7 ±0. 072vs 0. 675 7±0. 026. P 〈 0. 05 ; IL - 8 mRNA ： 0. 1541 ±0. 0. 019 vs 0. 208 1 ±0. 030. P 〈 0. 05 ）. Conclusion The selective MIF activity inhibitor, ISO - 1, can inhibit the invasion of Lovo cells, possibily by inhibition of tautomerase activities of MIF and down - regulation of MMP - 9 and IL - 8 levels.

关 键 词：大肠肿瘤 巨噬细胞移动抑制因子 互变异构酶活性 ISO-1 RT-PCR 

分 类 号：R734.2[医药卫生—肿瘤] R735.34[医药卫生—临床医学]