沉默单核细胞趋化蛋白-1基因降低人食管癌EC109细胞的迁移及侵袭能力  

Knock-down of Monocyte Chemoattractant Protein-1 Inhibits Migration and Invasion of Human Esophageal Carcinoma EC109 Cells

作　　者：张超贤[1] 韩宇[1] 常廷民[1] 李光艳[1] 

机构地区：[1]新乡医学院第一附属医院消化内科,河南卫辉453100

出　　处：《中国生物化学与分子生物学报》2016年第6期678-685,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：河南省教育厅科研基金项目(No.20073200143)资助~~

摘　　要：单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)是白色脂肪细胞分泌的炎症趋化刺激因子,属于趋化因子CC亚族,可促进肿瘤血管形成和细胞外基质降解,从而促进肿瘤细胞的浸润与转移。沉默MCP-1基因可显著抑制恶性肿瘤生长及转移,但其作用的分子机制尚不完全清楚。本研究应用小干扰RNA技术沉默人食管癌EC109细胞中MCP-1表达。细胞划痕试验显示,与对照组相比,沉默MCP-1基因可明显抑制食管癌EC109细胞迁移能力。Transwell侵袭实验显示,沉默MCP-1基因后,EC109细胞侵袭能力降低。Western印迹试验和RT-PCR试验揭示,沉默MCP-1基因后,细胞中MMP-7、MMP-9、TGF-β_1及VEGF表达水平显著下降。研究结果提示,沉默MCP-1基因可通过抑制MMP-7、MMP-9、TGF-β_1及VEGF表达,降低癌细胞迁移及侵袭能力。Monocyte chemoattractant protein-1（ MCP-1） is a member of the CC chemokine family,which is closely correlated to carcinoma cell migration and invasion. MCP-1 is highly expressed in human esophageal carcinoma EC109 cells. The migration and invasion abilities of human EC109 cells decreased after inhibition of MCP-1 expression. To explore the effects of small interfering RNA（ siRNA） silencing MCP-1 gene on migration and invasion of human EC109 cells,MCP-1-specific siRNA was designed and synthesized primarily. Human EC109 cells were transfected for 120 hours. The control groups were established accordingly by using siRNA with insignificant order and no transfection. The wound healing assay showed that silencing MCP-1 gene significantly inhibited the abilities of cells migration compared with all the control groups. The transwell experiment demonstrated that silencing MCP-1 gene led to decreased invasion abilities of EC109 cells. Furthermore,Western blotting and RT-PCR assay revealed that the expressions of MCP-1,MMP-7,MMP-9,TGF-β_1and VEGF were decreased after the silencing of MCP-1 gene,and there were obvious positive correlations among MCP-1,MMP-7,MMP-9,TGF-β_1and VEGF expressions. These data suggested that RNA interference of MCP-1 could inhibit carcinoma cell migration and invasion regulated by the expression of MCP-1,MMP-7,MMP-9,TGF-β_1and VEGF expressions.

关 键 词：小干扰RNA 单核细胞趋化蛋白-1 人食管癌EC109细胞 迁移及侵袭 

分 类 号：R735.1[医药卫生—肿瘤]