间质作用因子1通过正调控窖蛋白1抑制FBJ-S1-H的迁移性  

Stromal interaction molecule 1(Stim1) suppresses FBJ-S1-H cell migration through caveolin 1(Cav1)

作　　者：黄澄澄[1] 姜嘉慧[2] 吴英良[1] 山形达也[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [2]日本九州大学生物资源环境科学府,日本福冈8128581

出　　处：《沈阳药科大学学报》2011年第12期976-984,共9页Journal of Shenyang Pharmaceutical University

摘　　要：目的证明间质作用因子(stromal interaction molecule1,Stim1)在FBJ诱导的小鼠骨肉瘤细胞中的抑癌作用。方法在Stim1高表达的FBJ-S1-H细胞采用Stim1以siRNA干扰技术得到Stim1沉默的几株S1-H单克隆细胞株,通过细胞行为学方法和RT-PCR技术对其mRNA进行研究,通过明胶酶谱法对细胞基质金属酶活性进行研究。结果通过细胞行为学方法证明,Stim1的沉默提高了细胞的迁移性,通过对mRNA表达的研究发现,Stim1沉默引起了多种基因表达的变化,其中包括基质金属酶9(matrix mexalloprotelnase 9,MMP-9)的升高,窖蛋白(caveolinl,Cav1),甾醇调控因子Srebf1的降低等,提高单克隆细胞中的Cav1含量可以使细胞迁移性降低。结论实验结果证明在FBJ-S1-H细胞中,Stim1能够抑制细胞的移动性,沉默Stim1的表达能够提高细胞的迁移性。Objective To demonstrate the tumor suppressor function of Stiml in FBJ-S1-H cells. Methods Stiml was silenced in FBJ-S1-H cell, and three different monoclonal cell lines were obtained, namely E4,5 si and 14si. Cell motility and mRNA expression of these monoclonal cells were determined and enzyme activity was also examined using gelatin zymography. Results Wound healing assays indicated that monoclonal cells had high motility compared with parental cells and a scrambled siRNA transfected control monoclonal cell ScrB4. RT-PCR results showed a down regulation of Cavl and Srebfl expression, and a high MMP-9 expression in Stiml silenced monoclonal cells. Zymography results indicated the MMP-9 enzyme activity increased in parallel to mRNA expression in monoclonal cells. To investigate the relationship of Cavl, Srebfl and MMP-9 ,wound healing assay was performed. The recovery of Cavl expression by Cavl cDNA transfection in monoclone E4 showed suppressed cell motility, with a down-regulation of MMP-9 expression in mRNA level, while suppression of Srebfl in S1-H did not show any significant change in cell motility. Conclusions Judged from the suppressed cell motility, Stiml acts as a tumor suppressor gene in FBJ-S1-H cell. Silencing Stiml in FBJ-S1-H suppressed Cavl and elevated MMP-9 expression and is reflected in increased cell motility.

关 键 词：间质作用因子1 窖蛋白1 基质金属蛋白酶-9 甾醇调控因子1 伤痕愈合试验 肿瘤细胞的迁移性 

分 类 号：R963[医药卫生—微生物与生化药学]