摘要
选用高表达 α6β1并主要借其介导细胞在层粘连蛋白 ( laminin,LN)基质上粘附和铺展的肝癌细胞系 Bel740 2 ,采用“dominant negative”策略消除细胞表面 α6β1受体的功能 ,以研究 α6β1在人肝癌细胞生物学行为中的作用 .用缺失胞质功能域的 β4亚单位 c DNA( β4- tr)转染细胞 ,其表达产物β4- TR可以和α6高亲和性竞争性地结合 ,形成因β4- TR缺失胞质功能域而无生物学功能的α6β4- TR二聚体 ,取代α6β1 .结果显示在转染β4- tr的 Bel740 2细胞表面可以检测到大量的β4- TR,与内源性α6结合形成α6β4- TR二聚体 ,而细胞表面α6β1受体降至极低水平 .细胞在 LN基质上的铺展率降低 ,表明
An HCC cell line Bel7402,which expressed high level of integrin α6β1 on cell surface and applied mainly this integrin receptor to adhere and spread on LN,was identified to study the biological function of α6β1 in HCC.A dominant negative strategy was applied to eliminate the function of α6β1.The cytoplasmic functional domain truncated β4 subunit cDNA (β4 tr) was transfected to the cell.The expressive product β4 TR subunit bound preferentially with endogenous α6 subunit to form non functional α6β4 TR,substituting α6β1.Results showed that large amount of β4 TR was detected on the stable transfectants and formed α6β4 TR heterodimer,whereas the amount of α6β1 on cell surface decreased to very low level.The spreading of transfected cells on LN was significantly reduced,demonstrating that the function of α6β1 was eliminated.
出处
《中国生物化学与分子生物学报》
CSCD
2000年第1期133-137,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金!资助项目 ( 3 9780 0 3 8)
