【作者】 黎明；

【导师】 王娅兰；

【作者基本信息】 重庆医科大学 ， 病理学与病理生理学， 2008， 硕士

【摘要】 目的:探讨多聚(腺苷二磷酸核糖)聚合酶[poly(ADP-ribose) polymerase, PARP]抑制对小鼠结肠癌CT26细胞侵袭的影响及其可能的机制。方法:应用细胞-基质粘附试验、细胞运动及侵袭试验,观察PARP抑制剂5-氨基异喹啉酮(5-aminoisoquinolinone, 5-AIQ)抑制CT26细胞PARP前后,其基质粘附、运动及侵袭能力的变化;采用western blot检测5-AIQ对CT26细胞整合素β1(Integrinsβ1)、基质金属蛋白酶(matrix metalloproteinases, MMPs)MMP-2、MMP-9表达的影响,并用明胶酶谱法检测5-AIQ对明胶酶MMP-2和MMP-9活性的影响。结果:1.细胞-基质粘附实验结果显示,与未处理组比较,经5-AIQ处理过的CT26细胞与纤维连接蛋白的粘附率有明显降低(P<0.01);transwell运动实验结果表明,5-AIQ处理组迁移穿过微孔膜的细胞数较未处理组明显减少(P<0.01);同样transwell侵袭实验表明,5-AIQ处理组的CT26细胞侵袭穿透Matrigel胶,而到达膜下表面的细胞数较未处理组也明显减少(P<0.01).2.western blot结果提示,PARP抑制剂5-AIQ处理过的CT26细胞其整合素β1、MMP-2和MMP-9的蛋白表达较5-AIQ未处理的CT26细胞有明显减弱,差别均具有统计学意义(P值分别是:整合素β1 P<0.01、MMP-2 P<0.01、MMP-9 P<0.05);而用明胶酶谱法检测明胶酶MMP-2和MMP-9的活性时,在5-AIQ处理组和未处理组之间同样存在明显差异(P值分别是:MMP-2 P<0.05、MMP-9 P<0.01)。结论:1. PARP抑制剂5-AIQ可降低CT26细胞基质粘附、运动及侵袭能力,PARP在肿瘤侵袭中可能发挥作用。2. PARP抑制剂5-AIQ使CT26细胞整合素β1、MMP-2和MMP-9的蛋白表达降低,并可使MMP-2和MMP-9酶的活性抑制。提示可能PARP抑制,可能通过抑制肿瘤侵袭转移相关的因子整合素β1、MMP-2和MMP-9的表达和活性,从而影响CT26细胞的侵袭。更多还原

【Abstract】 Objective: The aim is to study the effect of poly(ADP-ribose) polymerase (PARP) inhibitor on the invasion of murine colon carcinoma CT26 cell lines in vitro,and its mechanism.Methods: CT26 cell lines were treated with or without PARP inhibitor 5-aminoisoquinolinone (5-AIQ) in vitro.The effects of 5-AIQ on the cell adhesion ,migration and invasion of CT26 were observed by cell- matrix adhesion assay, cell migration assay and matrigel invasion assay, respectively. The expressions of Integrinβ1, MMP-2 and MMP-9 of CT26 cells with or without 5-AIQ were investigated by Western Blot and the activities of MMP-2 and MMP-9 were detected by zymography.Results: 1. The result of cell-matrix adhesion assay showed that A values of 5-AIQ-treated groups were significantly reduced, when compared with untreated group (P<0.01). Meanwhile, comparing with untreated group, the migration and invasion potential of CT26 were both reduced by 5-AIQ treatment (P<0.01).2. The expressions of Integrinβ1, MMP-2 and MMP-9 of 5-AIQ-treated group were all significantly lower than the 5-AIQ-untreated group in western blot assay (P value were P<0.01, P<0.01, P<0.05, respectively). Interestingly, the activities of MMP-2 and MMP-9 were also attenuated in 5-AIQ-treated group comparing with 5-AIQ-untreated group (P value were P<0.05, P<0.01, respectively).Conclusion:1. The data suggest that PARP inhibitor 5-AIQ could reduce the cell-matrix adhesion, migration and invasion in CT26 cells. PARP inhibition might have a contribution for tumor invasion in some degree.2. 5-AIQ reduced the expressions of Integrinβ1, MMP-2 and MMP-9, and the activities of MMP-2 and MMP-9, which suggested that PARP inhibition suppressed the invasion-relative factors Integrinβ1, MMP-2 and MMP-9, then effected the metastasis of CT26 cells.更多还原