巨噬细胞游走抑制因子在卵巢癌侵袭中的作用与意义

Clinical significance of macrophage migration inhibitory factor in invasion of ovarian cancer

背景与目的:巨噬细胞游走抑制因子(macrophage migration inhibitory factor,MIF)与肿瘤的恶性进展密切相关。本研究探讨MIF基因对卵巢癌HO-8910和OVCAR-3细胞侵袭和增殖能力的影响及其在卵巢癌组织中表达的意义。方法:瞬时转染小干扰RNA(small interferingRNA,siRNA)靶向敲除MIF基因,RT-PCR和Westernblot检测MIF在mRNA和蛋白水平的表达,通过体外迁移、侵袭实验和噻唑蓝比色法(MTT)分析MIF对HO-8910和OVCAR-3细胞迁移、侵袭和增殖能力的影响;免疫组织化学检测MIF蛋白在卵巢癌组织中的表达情况。结果:与阴性对照组细胞比较,瞬时转染MIFsiRNA的HO-8910和OVCAR-3细胞中MIF基因表达水平明显降低。MTT结果显示,转染MIFsiRNA的两株卵巢癌细胞增殖率显著低于阴性对照组(P<0.05)。转染MIFsiRNA的HO-8910细胞穿膜细胞数(MIF-si1:48.0±7.3;MIF-si2:38.0±3.6)与阴性对照组(78.0±8.5)比较差异有统计学意义(P<0.05),其穿过铺了Matrigel膜的细胞数(MIF-si1:35.0±5.0;MIF-si2:30.0±5.6)也显著少于阴性对照组(P<0.05)。同样,转染了MIFsiRNA的OVCAR-3细胞穿膜细胞数(MIF-si1:40.0±4.5;MIF-si2:42.0±3.0)与阴性对照组(65±2.1)比较,差异也有统计学意义(P<0.05),其穿过铺了Matrigel膜的细胞数(MIF-si1:25.0±3.0;MIF-si2:27.0±3.4)也明显低于阴性对照组(P<0.05)。53.6%(37/69)卵巢癌组织中有MIF蛋白表达,MIF蛋白表达与肿瘤临床分期呈显著正相关(P<0.01)。结论:MIF基因可能通过促进肿瘤细胞的增殖和侵袭能力在卵巢癌的发生发展中发挥重要作用,有可能成为分子靶向治疗卵巢癌的潜在靶点。

Background and Objective. Macrophage migration inhibitory factor （MIF） is closely related to tumorigenesis. This study was to investigate the effects of MIF gene on migration, invasion and proliferation of ovarian cancer cells and to evaluate the significance of MIF protein expression in ovarian cancer tissues. Methods. Small interfering RNA was used to transiently knock down the expression of MIF gene in HO-8910 and OVCAR-3 cells. The effect of RNAi was assessed by RT-PCR and western blot. The migration, invasion and proliferation of ovarian cancer cells were determined by transwell chamber assay, invasion assay and MTT assay, respectively. Immunohistochemistry was utilized to examine the expression status of MIF in ovarian cancer tissues. Results. MIF RNAi significantly inhibited MIF expression in HO-8910 and OVCAR-3 cells and decreased cell proliferation of the two cells （P〈0.05）. The numbers of migrated and invaded HO-8910 ceils were significantly less in the MIF-si1 and MIF-si2 groups than in the NC group, respectively [migration, （48.0±7.3） and （38.0±3.6） vs. （78.0±8.5）, P〈0.05; invasion. （35.0±5.0） and （30.0±5.6） vs. （65.0±4.6）, P〈0.05]. The numbers of migrated and invaded OVCAR-3 cells were significantly less in the MIF siRNA groups than in the NC group, respectively [migration. （40.0± 4.5） and （42.0±3.0） vs. （65±2.1）, P〈0.05;invasion. （25.0±3.0） and （27.0±3.4） vs. （48.0±2.4）, P〈0.05]. Positive expression of MIF protein was detected in 53.5% of ovarian carcinoma tissues and was positively correlated to clinical stages of patients （P〈0.01）. Conclusion. MIF might play an important role in the pathogenesis and progression of ovarian cancer. Thus, MIF might be used as a potential therapeutic target in ovarian cancer.