HAI-1过表达对SW620细胞体外生长和运动能力的影响

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HAL-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。

EFFECT OF HAI-1 OVER-EXPRESSION ON IN VITRO GROWTH AND MIGRATION/MOTILITY OF SW620 CELLS

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 cD- NA of human HAI-1 into mammalian expression vector pcDNA3.1 ( ), and transfected the con- struct 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 evalu- ate 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 differ- ences of growth of HAI-1-transfected cells when compared with the control groups. Trans-mem- brane migration assay and wound-healing motility assay showed significant inhibition of cell mi- gration/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.