巨噬细胞移动抑制因子在肾癌中的表达及对肾癌细胞侵袭能力的影响

摘要

Objective To explore the expression of macrophage migration inhibitory factor (MIF) in renal carcinoma tissue and its effect on invasion of renal carcinoma cell.Methods Forty-eight cases of patients definitely diagnosed with renal cancer in the Department of Pathology,the Inner Mongolia Autonomous Region People's Hospital and Baotou Municipal Tumor Hospital from March 2013 to March 2015 served as the research subjects.The expressions of MIF protein and mRNA in renal carcinoma tissues and adjacent normal tissues,renal carcinoma cell lines and normal renal tubular epithelial cells HK-2 were detected by adopting the Western blot and RT-PCR.siRNA MIF and siRNA NC were transfected into renal carcinoma cells by liposome.The transfection effect was detected by Western blot and RT-PCR.The cell viability and invasion ability were measured by MTT and Transwell method respectively.The expression of hypoxia inducible factor 1-alpha (HIF-1α) and matrix metalloproteinase 9 (MMP9) was measured by Western blot.Results The expression levels of MIF protein and mRNA in renal carcinoma tissues were significantly higher than those in adjacent normal tissues (0.89±0.09 vs.0.18±0.02,1.19±0.04 vs.0.19±0.01,P＜0.05).The expression levels of MIF protein and mRNA in renal cancer cell line ACHN,769-p,786-O,Caki-1 cells were higher than those in renal tubular epithelial HK-2 cells (P＜0.05).Compared with the siRNA NC group,the expression levels of MIF protein and mRNA in siRNA MIF group were significantly down-regulated (P＜0.05),the cell viability and invasion ability were reduced (P＜0.05),and the expressions of HIF-1α and MMP9 were down regulated (P＜0.05).Conclusion MIF is highly expressed in renal carcinoma,downregulating MIF expression can significantly inhibit the renal cancer cell invasion,which might be related with inhibiting HIF-1α/MMP9 signaling pathway.%目的 探讨巨噬细胞移动抑制因子(MIF)在肾癌组织中的表达及对肾癌细胞侵袭能力的影响.方法 以内蒙古自治区人民医院和包头市肿瘤医院2013年3月至2015年3月病理室确诊的49例肾癌患者为研究对象,采用蛋白质印迹法(West-ern blot)及反转录(RT)-PCR法检测肾癌组织及癌旁正常组织、肾癌细胞株及正常肾小管上皮细胞HK 2中MIF蛋白及mRNA表达;利用脂质体将siRNA MIF及siRNA NC转染到肾癌细胞中,Western blot及RT-PCR检测转染效果,噻唑蓝(MTT)法检测细胞活力,Transwell法检测细胞侵袭能力,Western blot检测细胞中缺氧诱导因子-1α(HIF-1α)及基质金属蛋白酶9(MMP9)的表达.结果 肾癌组织中MIF蛋白及mRNA表达水平明显高于癌旁正常组织(0.89±0.09 vs.0.18±0.02,1.19±0.04 vs.0.19±0.01,P＜0.05),肾癌细胞株ACHN、769-p、786-O、Caki-1中MIF蛋白及mRNA表达水平明显高于肾小管上皮细胞HK-2(P＜0.05).与siRNA NC组比较,siRNA MIF组中MIF蛋白及mRNA表达水平明显下调(P＜0.05),细胞活力及侵袭能力降低(P＜0.05),HIF-1α及MMP9表达水平下调(P＜0.05).结论 MIF在肾癌中高表达,下调MIF表达能明显抑制肾癌细胞的侵袭能力,可能与抑制HIF-1α/MMP9信号通路有关.