下调星形细胞上调基因1表达可抑制膀胱癌T24细胞的增殖和侵袭

Down-regulation of astrocyte elevated gene-1 expression inhibits the proliferation and invasion of human bladder cancer T24 cells

目的 :检测星形细胞上调基因1(astrocyte elevated gene-1,AEG-1)在人膀胱癌细胞中的表达,探讨AEG-1表达下调对膀胱癌T24细胞增殖和侵袭能力的影响。方法 :采用蛋白质印迹法和免疫荧光染色法检测正常膀胱尿路上皮细胞SV-HUC-1以及膀胱癌细胞株RT4、J82、5637和T24中AEG-1蛋白的表达水平。将特异性靶向AEG-1基因的sh RNA重组病毒感染自身高表达AEG-1的膀胱癌T24细胞,经嘌呤霉素筛选获得稳定感染细胞系,采用蛋白质印迹法验证AEG-1基因被沉默的效果;然后,采用CCK-8法、划痕愈合实验和Transwell小室法分别检测AEG-1基因沉默对膀胱癌T24细胞增殖、迁移和侵袭能力的影响。结果 :4种膀胱癌细胞中AEG-1蛋白表达水平明显高于正常膀胱尿路上皮细胞(P值均<0.05)。AEG-1 sh RNA重组病毒感染能明显下调膀胱癌T24细胞中AEG-1蛋白的表达(P<0.05)。与感染阴性对照sh RNA重组病毒的T24细胞相比,AEG-1 sh RNA下调AEG-1表达后T24细胞的增殖受到明显抑制(P<0.05),而且细胞的迁移和侵袭能力均明显减弱(P值均<0.05)。结论 :沉默AEG-1基因表达可抑制膀胱癌细胞的增殖、迁移及侵袭,提示AEG-1基因可能具有促进膀胱癌细胞发生恶性生物学行为的作用。

Objective： To detect the expression level of astrocyte elevated gene-1 （AEG-1） in human bladder cancer cell lines, and to investigate the effects of down-regulation of AEG-1 expression on proliferation and invasion of bladder cancer T24 cells. Methods： The expression level of AEG-1 protein in normal bladder urinary epithelial cell line SV-HUC-1 and bladder cancer cell lines RT4, J82, 5637 and T24 was detected by Western blotting and immunofluorescence staining. The recombinant virus with specific shRNA targeting AEG-1 gene was infected into bladder cancer T24 cells with high-expression of endogenous AEG-1. The stable infected clones were screened by puromycin, and the silencing efficiency ofAEG-1 gene was confirmed by Western blotting. Then the effects of AEG-1 gene-silencing on the cell proliferation, migration and invasion were examined by CCK-8 method, wound healing assay and Transwell chamber assay, respectively. Results： The expression levels of AEG-1 in 4 bladder cancer cell lines were significantly higher than that in normal bladder urinary epithelial cell line （all P 〈 0.05）. After infection AEG-1 shRNA with the recombinant virus carrying AEG-1 shRNA, the expression level of AEG-1 protein was effectively down-regulated in bladder cancer T24 cells （P 〈 0.05）. As compared with the negative shRNA transfection group, the proliferation of T24 cells after AEG-1 gene- silencing was obviously inhibited （P 〈 0.05）, and the migration and invasion abilities of T24 ceils after AEG-1 gene-silencing were significantly decreased （both P 〈 0.05）. Conclusion： SilencingAEG-1 gene expression can inhibit the proliferation, migration and invasion of bladder cancer cells, which suggests that AEG-1 maybe paly a role in promoting the malignant biological behavior of bladder cancer cells.