整合素α6β1增强人肝癌细胞侵袭性机制的探讨

目的 研究层粘连蛋白受体α６β１整合素在人肝癌细胞侵行为中的作用。方法 采用ｄｏｍｉｎａｎｔ ｎｅｇａｔｉｖｅ策略,以无功能的α６β４－ＴＲ代替细胞表面α６β１受体,从而消除了人肝癌细胞Ｂｅｌ－７０４２ α６β１受体的功能。用划痕法和Ｂｏｒｄｅｎ小室法分别检测细胞的迁移和侵袭,明胶酶谱分析法检测细胞基质金属蛋白ｍａｔｒｉｘ ｍｅｔａｌｌｏｐｒｏｖｔｅｉｎａｓｅｓ,ＭＭＰａ）的分泌。结果 消除α６β

Mechanism of enhanced invasiveness of human hepatocellular carcinoma by integrin alpha 6 beta 1]

OBJECTIVE: To study the mechanism of enhanced invasiveness of human hepatocellular carcinoma (HCC) by laminin (LN) receptor integrin alpha 6 beta 1. METHODS: A stable transfectant of HCC Bel 7402 cell line, the alpha 6 beta 1 receptors of which were replaced by non-functional LN receptor alpha 6 beta 4-TR using dominant negative strategy, was used. Motility of the non-transfected, mock transfected and transfected tumor cells was assessed by the number of cells migrated to an area scratched out of cells on tumor cell monolayer. Tumor cell invasion was examined by cell penetration through a Matrigel layer in Boyden chamber. Matrix metalloproteinases (MMPs) secretion was detected by gelatin zymography. RESULTS: The migration of HCC Bel 7402 cells expressing dominant negative alpha 6 beta 1 was significantly decreased. Their invasive capability was decreased by 46.7%. The secretion of MMP-9 was almost totally inhibited and the activated MMP-2 was decreased by 43.9%. CONCLUSION: The decrease in cell motility, invasiveness and MMPs secretion of HCC cells expressing dominant negative alpha 6 beta 4 TR implies that LN-integrin interaction plays important role in HCC progression.