下调窖蛋白-1表达抑制小鼠肝癌H22细胞侵袭能力  

Knockdown of Caveolin-1 by siRNA Inhibits the Invasive Capability of Mouse Hepatoma H22 Cell

作　　者：汪淑晶[1] 樊建慧[1] 于生金[2] 张嘉宁[1] 

机构地区：[1]大连医科大学生物化学与分子生物学教研室糖生物学研究所,辽宁省大连116044 [2]辽东学院基础医学院,辽宁省丹东118000

出　　处：《中国生物化学与分子生物学报》2011年第4期323-329,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目(No.31000372;No.30970648)~~

摘　　要：窖蛋白-1在不同肿瘤中发挥作用不同.本研究以小鼠肝癌细胞H22为研究对象,观察下调窖蛋白-1表达对H22细胞侵袭能力的影响,并探讨其可能的分子机制.利用RT-PCR和Western印迹法检测了窖蛋白-1在H22及小鼠正常肝细胞IAR20中的表达.结果显示,窖蛋白-1在H22中的表达高于其在IAR20中的表达,提示窖蛋白-1高表达可能与H22细胞恶性表型有关.RNA干扰和凝集素印记实验结果显示,窖蛋白-1-siRNA能够有效抑制窖蛋白-1mRNA和蛋白表达,并抑制细胞表面N-聚糖β1,6GlcNAc分支形成.Transwell细胞迁移和侵袭实验结果显示,与未转染组和siRNA对照组比较,转染窖蛋白-1-siRNA的H22细胞迁移和侵袭数目明显减少.本研究证明,下调窖蛋白-1表达可抑制H22细胞表面N-聚糖β1,6GlcNAc分支形成,从而抑制细胞迁移和侵袭能力.Caveolin-1 is a major structural protein of caveolae and plays important roles in signal transduction and tumorigenesis.Caveolin-1 gene was regarded as both a tumor suppressor gene and an oncogene.Our previous study demonstrated that caveolin-1 was highly expressed in mouse hepatoma cell lines with lymphatic metastasis potential.However,the role of caveolin-1 in cellular migration and invasion remains unknown.Here,we used RNA interference to study the effects of silencing caveolin-1 expression on the invasive ability of mouse hepatoma H22 cells.Via targeting the mRNA region of caveolin-1,siRNA effectively suppressed caveolin-1 mRNA and protein levels.This resulted in the decreased invasive ability of H22 cells in vitro.In addition,down regulation of caveolin-1 expression suppressed the formation of N-glycan β1,6 GlcNAc branching on H22 cell surface.These results indicate that caveolin-1 could play an active role in mediating the migration and invasion of mouse hepatoma cells and might be a potential target for gene and antitumor drug therapy.

关 键 词：窖蛋白-1 肝癌 侵袭 糖基化 

分 类 号：Q73[生物学—分子生物学]