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HAI-1过表达对SW620细胞体外生长和运动能力的影响
引用本文:曹江,叶景佳.HAI-1过表达对SW620细胞体外生长和运动能力的影响[J].实验生物学报,2005,38(3):183-189.
作者姓名:曹江  叶景佳
作者单位:[1]浙江大学邵逸夫临床医学研究所,杭州310016 [2]浙江大学肿瘤研究所,杭州310009
摘    要:肝细胞生长因子激活因子抑制因子1(hepatocyte growth factor activator inhibitor type 1,HAI-1)能有效抑制肝细胞生长因子激活因子(hepatocyte growth factor activator,HGFA)和丝氨酸蛋白酶Matriptase的活性,并可通过对HGFA和Matriptase活性的调控参与HGF/c—Met信号传导途径。为了解HAI-1在肿瘤细胞的生长和运动中的作用,本研究将人HAI-1基因全长cDNA克隆至pcDNA3.1( )表达载体,并转染人肠癌SW620细胞,用Western blot验证了转染细胞中HAI-1的表达情况,并分别利用生长曲线、软琼脂集落形成、穿膜运动和扩散运动测定等方法检测了HAI-1过表达对SW620细胞生长和运动能力的影响。生长曲线和软琼脂集落形成测定都显示出HAI-1转染细胞与对照组相比差异不十分明显。穿膜运动和扩散运动测定则均显示了HAI-1过表达对细胞运动能力有明显的抑制。因此,HAI-1的过表达虽然在体外对肿瘤细胞生长影响较小,但可以抑制肿瘤细胞的运动迁移能力。

关 键 词:运动能力  过表达  体外生长  肝细胞生长因子  全长cDNA克隆  Western  丝氨酸蛋白酶  信号传导途径  肿瘤细胞  pcDNA3  激活因子  转染细胞  生长曲线  扩散运动  抑制因子  type  表达载体  blot  生长影响  迁移能力  Met  HGF  测定  活性

Effect of HAI-1 over-expression on in vitro growth and migration/motility of SW620 cells]
Jiang Cao,Jing Jia Ye.Effect of HAI-1 over-expression on in vitro growth and migration/motility of SW620 cells][J].Acta Biologiae Experimentalis Sinica,2005,38(3):183-189.
Authors:Jiang Cao  Jing Jia Ye
Institution:Sir Run Run Shaw Clinical Research Institute, Zhejiang University, Hangzhou 310016, China. caoj@zju.edu.cn
Abstract:Hepatocyte growth factor activator inhibitor type 1 (HAI-1) is an effective inhibitor for hepatocyte growth factor activator (HGFA) and serine protease Matriptase, and is involved in HGF/c-Met signaling pathway by regulating activities of HGFA and Matriptase. In an attempt to elucidate the roles HAI-1 play in tumor cell growth and migration, we subcloned full length cDNA of human HAI-1 into mammalian expression vector pcDNA3.1 (+), and transfected the construct into human colorectal cancer cell SW620. The expression of HAI-1 in transfected SW620 cells was demonstrated by Western blot. Growth curve determination, soft-agar colony formation assay, trans-membrane migration assay and wound-healing cell motility assay were used to evaluate the effects of over-expression of HAI-1 on cell growth and migration/motility. Both of the growth curve and soft-agar colony formation assays showed that there were no significant differences of growth of HAI-1-transfected cells when compared with the control groups. Trans-membrane migration assay and wound-healing motility assay showed significant inhibition of cell migration/motility ability by the over-expressed HAI-1. Our data suggest that over-expression of HAI-1 in SW620 cells has weak effect on cell growth in vitro, but can significantly inhibit cell migration/motility.
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