Effects of over-expression of macrophage migration inhibitory factor on the expression of interleukin-8 and martix metalloproteinase-9 of human cervical cancer SiHa cells
10.3760/cma.j.issn.1006-9801.2013.10.001
- VernacularTitle:过表达巨噬细胞移动抑制因子对子宫颈癌SiHa细胞中白细胞介素8及基质金属蛋白酶9表达的影响
- Author:
Hongxia GUO
;
Suhui WU
;
Rui JIA
;
Haixia SHANG
- Publication Type:Journal Article
- Keywords:
Macrophage migration inhibitory factor;
Inrerleukin-8;
Martix metalloproteinase-9;
Transfection;
Uterine cervical neoplasms
- From:
Cancer Research and Clinic
2013;25(10):649-653,658
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of macrophage migration inhibitory factor (MIF) overexpression on the expression of interleukin-8 (IL-8),martix metalloproteinase-9 (MMP-9) and invasion of human cervical cancer SiHa cells.Methods Chemical synthesis MIF eDNA gene,designed primer sequence including XhoI and BamHI enzyme sites,MIF gene was amplified by polymerase chain reaction (PCR),constructed eukaryotic expression vector pEGFP-N1/MIF and transfected into SiHa cells using Lipofectamine and won over-expression of MIF.The expression of MIF in supernatant fluid was detected by ELISA,the expression of MIF,IL-8,MMP-9 in both mRNA and protein levels were detected by real-time fluorescence quantitative-PCR and immunocytochemistry respectively.The effect of over-expressed MIF on migration was detected by Boyden small chamber.Results The expression of protein in supernatant fluid transfected with pEGFP-N1/MIF was significantly increased (Fgroup =8267.564,P ＜ 0.01),the expression of MIF,IL-8,MMP-9 in both mRNA and protein in SiHa cells transfected with pEGFP-N1/MIF were significantly increased (F values were 7019.619,2148.094,3303.540,1565.114,2807.300,523.466,P ＜ 0.01),and there was a positive correlation among MIF,IL-8,MMP-9 expression in both mRNA and protein (r values were 0.865,0.895,0.934,0.908,P ＜ 0.01).Invasion ability in SiHa cells transfected with pEGFP-N1/MIF was obviously increased (F=3430.898,P＜ 0.01).Conclusion The over-expression MIF gene in SiHa cells can promote cervical cancer cell invasion and metastasis of ability,which could be associated with the upregulation of IL-8 and MMP-9 expression.
