ANTXR1对人食管癌ECA109细胞株迁移和侵袭能力的影响

Effects of ANTXR1 Gene on Migration and Invasion of Human Esophageal Cancer ECA109 Cells

目的研究炭疽毒素受体1(anthrax toxin receptor 1,ANTXR1)对食管癌细胞系ECA109的迁移及侵袭能力的影响及机制。方法采用化学合成ANTXR1基因的小干扰RNA(small interfering RNA,si RNA)下调该基因表达,采用实时荧光定量聚合酶链式反应(real-time polymerase chain reaction,RT-PCR)和Western blot检测转染后的细胞内ANTXR1、基质金属蛋白酶2(matirx metallo-proteinases 2,MMP 2)和基质金属蛋白酶9(matirx metallo-proteinases 9,MMP9)的表达,通过细胞划痕实验和Transwell侵袭实验分别检测ANTXR1对ECA109细胞迁移和侵袭的影响。结果将ANTXR1转染入ECA109后,细胞中ANTXR1 m RNA和蛋白水平都明显降低,MMP2、MMP9表达明显下调,ECA109细胞的迁移和侵袭能力显著降低,与对照组相比差异有统计学意义(P<0.01)。结论在食管癌细胞系ECA109中ANTXR1与细胞的迁移和侵袭能力相关,ANTXR1可能通过影响MMP2和MMP9的表达参与其中,有望成为食管癌早期诊断、分子治疗的靶点。

Objective To investigate the effects of anthrax toxin receptor 1 （ANTXR1） gene on migration and in- vasion of human esophageal cancer ECA109 cells and its possible mechanism. Methods Chemically synthesized small in- terfering RNA （siRNA） targeting ANTXR1 gene was transfected into ECA109 cells with LipofectamineTM 2000. The ex- pressions of ANTXR1 ,matrix metallo-proteinase 2 （MMP 2） and MMP 9 after transfection were detected by real-time polymerase chain reaction （RT-PCR） and Western blot. The effects of migration and invasion of ANTXR1 silencing on ECA109 cells were evaluated by scratch test and matrigel invasion assay. Results The ANTXR1 mRNA and protein lev- els were significantly down-regulated in ECA109 cells transfected with ANTXRl-targeting siRNA2 as compared with con- trol group （P〈0. 01）. The expressions of MMP2 and MMPg, migration and invasion of ECA109 cells were lower than those in control group （P〈0. 01）. Conclusion ANTXR1 gene in esophagus cancer is related to migration and invasion of ECA109 cells by influencing MMP2 and MMP9 expressions. It is expected to be a target for early diagnosis and molecular therapy of esophageal squamous cell carcinoma.