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目的探讨腺苷酸环化酶相关蛋白1(CAP1)对人脑胶质瘤细胞株U87、U251迁移、侵袭和黏附的影响及其机制。方法运用小干扰RNA(siRNA)技术分别转染U87、U251细胞,Western blot检测CAP1、MMP-2、MMP-9、黏着斑激酶(FAK)和含有酪氨酸激酶活性的Src的表达。应用细胞划痕实验及Transwell迁移、侵袭实验检测U87、U251细胞的迁移和侵袭能力。应用细胞黏附实验检测细胞黏附能力。结果与阴性对照组比较,CAP1-siRNA组U87、U251细胞中CAP1蛋白表达量降低,MMP-2、MMP-9、FAK和Src的蛋白表达量均减少(P<0.05)。CAP1-siRNA组较阴性对照组细胞的迁移能力降低(P<0.05)。沉默CAP1后,U87、U251细胞的迁移、侵袭能力下降(P<0.05)。CAP1-siRNA组黏附细胞个数少于阴性对照组(P<0.05)。结论 CAP1-siRNA靶向干扰了CAP1的表达,同时下调MMP-2、MMP-9、FAK和Src的表达。下调CAP1的表达水平能够显著降低人脑胶质瘤细胞株U87、U251迁移、侵袭和黏附能力。
Objective To investigate the effect of adenylate cyclase-associated protein 1 (CAP1) on the migration, invasion and adhesion of human glioma cell lines U87 and U251 and its mechanism. Methods U87 and U251 cells were transfected with small interfering RNA (siRNA). The expression of CAP1, MMP-2, MMP-9, FAK and Src with tyrosine kinase activity were detected by Western blot. Cell migration assay and Transwell migration assay were used to detect the migration and invasion of U87 and U251 cells. Cell adhesion assay was used to detect cell adhesion. Results Compared with the negative control group, the expression of CAP1 in U87 and U251 cells of CAP1-siRNA group decreased and the protein expression of MMP-2, MMP-9, FAK and Src decreased (P <0.05). The migration ability of CAP1-siRNA group was lower than that of negative control group (P <0.05). After silencing CAP1, the migration and invasion ability of U87 and U251 cells decreased (P <0.05). The number of adherent cells in CAP1-siRNA group was less than that in negative control group (P <0.05). Conclusions CAP1-siRNA targeting interfered with the expression of CAP1 and down-regulated the expressions of MMP-2, MMP-9, FAK and Src. Down-regulation of CAP1 expression could significantly reduce the migration, invasion and adhesion of human glioma cell lines U87 and U251.