【摘要】 目的:探讨氯通道ClC-2反义寡核苷酸(AON)抑制ClC-2基因表达对顺铂(DDP)作用下神经胶质瘤U251细胞侵袭力的影响,为ClC-2成为神经胶质瘤综合治疗的新基因靶点提供理论依据。方法:U251细胞按处理方法不同分为对照组(转染无义寡核苷酸)、AON组(转染ClC-2反义寡核苷酸)、DDP组(无义寡核苷酸与顺铂联用)和AON+DDP组(ClC-2反义寡核苷酸与顺铂联用)。应用RT-PCR检测相关基因ClC-2、cyclin D1、MMP-2、MMP-9 mRNA表达;Transwell小室侵袭实验检测各组细胞侵袭力;Transwell小室迁移实验检测各组细胞迁移率。结果:与对照组比较,AON组、DDP组和AON+DDP组ClC-2、cyclin D1、MMP-2、MMP-9 mRNA表达率降低(P<0.05);Transwell小室侵袭实验中对照组、AON组、DDP组和AON+DDP组侵袭细胞数分别为81.00±4.58、42.01±4.36、48.33±2.52和12.67±5.13;与对照组比较,AON组、DDP组和AON+DDP组穿透细胞数明显降低(P<0.05)。Transwell小室迁移实验中AON组、DDP组、AON+DDP组细胞迁移率(%)分别为72.40±2.20、51.48±1.29、37.87±1.50,与对照组比较均明显降低(P<0.05)。结论:①转染ClC-2反义寡核苷酸能有效抑制ClC-2 mRNA表达;②抑制ClC-2 mRNA能降低U251细胞的迁移力和侵袭力;③ClC-2 mRNA表达降低可促进顺铂对U251细胞迁移力和侵袭力的抑制作用。更多还原

【Abstract】 Objective To investigate the effect of chloride channel ClC-2 antisense oligonucleotide(AON) on the cell invasion suppression of human glioma U251 cells induced by cisplatin(DDP).Methods The experiment was divided into four groups: control group(nonsense oligonucleotide),AON group(ClC-2 antisense oligonucleotide),DDP group(cisplatin+nonsense oligonucleotide),DDP+AON group(cisplatin+ClC-2 antisense oligonucleotide).The mRNA expressions of ClC-2,cyclin D1,MMP-2 and MMP-9 of U251 cells were detected by RT-PCR.The invasion of U251 was measured by Transwell assay.Results Compared with control group,the mRNA expression rates of ClC-2,cyclin D1,MMP-2,MMP-9 in AON group,DDP group,DDP+AON group were decreased(P<0.05).The numbers of invasion cells in control group,AON group,DDP group and AON +DDP group were 81.00±4.58,42.01±4.36,48.33±2.52 and 12.67±5.13,respectively.Compared with control group,the numbers of invasion cells in AON group,DDP group and AON +DDP group were significantly decreased(P<0.05).Compared with control group,the rates of cell migration(%) in AON group,DDP group and AON +DDP group were 72.40±2.20,51.48±1.29 and 37.87±1.50,respectively.Cell migration of U251 cells was remarkably decreased after infected with ClC-2 antisense and treated with cisplatin(P<0.05).Conclusion ①ClC-2 antisense oligonucleotide can inhibit the expression of ClC-2 mRNA in U251 cells.Inhibition of ClC-2 mRNA can suppress the mobility and invasion of U251 cells.③ClC2 antisense oligonucleotide can facilitate invasion suppression of U251 cells induced by DDP.更多还原