[Effect of HAI-1 over-expression on in vitro growth and migration/motility of SW620 cells].

Shi yan sheng wu xue bao Pub Date : 2005-06-01
Jiang Cao, Jing Jia Ye
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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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HAI-1过表达对SW620细胞体外生长和迁移/运动的影响
肝细胞生长因子激活因子抑制剂1型(HAI-1)是肝细胞生长因子激活因子(HGFA)和丝氨酸蛋白酶基质酶(Matriptase)的有效抑制剂,通过调节HGFA和基质酶的活性参与HGF/c-Met信号通路。为了阐明HAI-1在肿瘤细胞生长和迁移中的作用,我们将人HAI-1全长cDNA亚克隆到哺乳动物表达载体pcDNA3.1(+)中,并将其转染到人结直肠癌细胞SW620中。Western blot检测转染SW620细胞中HAI-1的表达。采用生长曲线测定、软琼脂集落形成试验、跨膜迁移试验和创面愈合细胞运动试验评价过表达HAI-1对细胞生长和迁移/运动的影响。生长曲线和软琼脂集落形成实验均显示,转染hai -1的细胞与对照组相比生长无显著差异。跨膜迁移实验和创面愈合运动实验显示,过表达的HAI-1显著抑制细胞迁移/运动能力。我们的数据表明,在SW620细胞中过表达HAI-1对细胞体外生长的影响较弱,但可以显著抑制细胞的迁移/运动性。
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