中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2011年
6期
456-460
,共5页
刘海燕%张新颖%王立俊%孙若鹏
劉海燕%張新穎%王立俊%孫若鵬
류해연%장신영%왕립준%손약붕
神经母细胞瘤%RNA干扰%肿瘤侵润%基质金属蛋白酶-9
神經母細胞瘤%RNA榦擾%腫瘤侵潤%基質金屬蛋白酶-9
신경모세포류%RNA간우%종류침윤%기질금속단백매-9
Neuroblastoma%RNA interference%Neoplasm invasiveness%Matrix metalloproteinase-9
目的 观察下调Astrocyte elevated gene-1(AEG-1)基因对神经母细胞瘤细胞中基质金属蛋白酶-9(MMP-9)表达以及细胞侵袭能力的影响.方法 根据RNA干扰原理,设计合成AEG-1 siRNA.通过荧光定量RT-PCR和Western blotting观察神经母细胞瘤细胞中AEG-1基因mRNA和蛋白下调情况;采用Western blotting观察下调AEG-1表达对神经母细胞瘤细胞中MMP-9蛋白表达的影响;通过侵袭实验(Transwell法)观察下调AEG-1表达对神经母细胞瘤细胞侵袭能力的影响.结论 采用AEG-1 siRNA转染神经母细胞瘤细胞,下调细胞内AEG-1的mRNA和蛋白表达(P＜0.05)后MMP-9蛋白表达抑制率分别为63.1%和58.9%.此外下调AEG-1表达使细胞侵袭能力明显减弱(P＜0.05).结论 AEG-1基因下调使神经母细胞瘤细胞中MMP-9的蛋白表达明显减少、活性明显减弱,同时使细胞的侵袭能力降低.由此可见AEG-1在肿瘤细胞的侵袭转移中可能扮演重要角色,将会成为神经母细胞瘤基因治疗的新靶点.
目的 觀察下調Astrocyte elevated gene-1(AEG-1)基因對神經母細胞瘤細胞中基質金屬蛋白酶-9(MMP-9)錶達以及細胞侵襲能力的影響.方法 根據RNA榦擾原理,設計閤成AEG-1 siRNA.通過熒光定量RT-PCR和Western blotting觀察神經母細胞瘤細胞中AEG-1基因mRNA和蛋白下調情況;採用Western blotting觀察下調AEG-1錶達對神經母細胞瘤細胞中MMP-9蛋白錶達的影響;通過侵襲實驗(Transwell法)觀察下調AEG-1錶達對神經母細胞瘤細胞侵襲能力的影響.結論 採用AEG-1 siRNA轉染神經母細胞瘤細胞,下調細胞內AEG-1的mRNA和蛋白錶達(P＜0.05)後MMP-9蛋白錶達抑製率分彆為63.1%和58.9%.此外下調AEG-1錶達使細胞侵襲能力明顯減弱(P＜0.05).結論 AEG-1基因下調使神經母細胞瘤細胞中MMP-9的蛋白錶達明顯減少、活性明顯減弱,同時使細胞的侵襲能力降低.由此可見AEG-1在腫瘤細胞的侵襲轉移中可能扮縯重要角色,將會成為神經母細胞瘤基因治療的新靶點.
목적 관찰하조Astrocyte elevated gene-1(AEG-1)기인대신경모세포류세포중기질금속단백매-9(MMP-9)표체이급세포침습능력적영향.방법 근거RNA간우원리,설계합성AEG-1 siRNA.통과형광정량RT-PCR화Western blotting관찰신경모세포류세포중AEG-1기인mRNA화단백하조정황;채용Western blotting관찰하조AEG-1표체대신경모세포류세포중MMP-9단백표체적영향;통과침습실험(Transwell법)관찰하조AEG-1표체대신경모세포류세포침습능력적영향.결론 채용AEG-1 siRNA전염신경모세포류세포,하조세포내AEG-1적mRNA화단백표체(P＜0.05)후MMP-9단백표체억제솔분별위63.1%화58.9%.차외하조AEG-1표체사세포침습능력명현감약(P＜0.05).결론 AEG-1기인하조사신경모세포류세포중MMP-9적단백표체명현감소、활성명현감약,동시사세포적침습능력강저.유차가견AEG-1재종류세포적침습전이중가능분연중요각색,장회성위신경모세포류기인치료적신파점.
Objective To investigate the effect of astrocyte elevated gene-1 (AEG-1)siRNA induced inhibition of invasion in neuroblastoma cells.Methods A small interference RNA(siRNA) targeting to AEG-1 mRNA(AEG-1 siRNA) was constructed and transfected into neuroblastoma cells with Lipofectamine 2000.A non-specific siRNA(control siRNA) and non-treatment were used as negative control group and blank group.The expression of AEG-1 mRNA was detected by RT-PCR.The proteins of AEG-1 and MMP-9 were detected by Western blotting.Cell invasion after AEG-1 knockdown was observed by Transwell assay.Results Compared with the control group,the expression of AEG-1 mRNA and protein were significantly decreased in the cells transfected with AEG-1 siRNAs(P＜0.05).AEG-1 knockdown by siRNA markedly decreased the expression of MMP-9 protein by 63.1% in neuroblastoma cells and the ability of cell invasion (P＜0.05).Conclusions The expression of AEG-1 mRNA is down-regulated by AEG-1 siRNA in neuroblastoma cells.Moreover,knockdown of AEG-1 expression in human neuroblastoma cells significantly inhibits the expression of MMP-9 protein and cell invasion.Therefore,AEG-1 may play a key role for MMP-9 activity and cell invasion,making it a potential target gene for gene-therapy in neuroblastoma.
