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

发布时间：2018-06-03 01:09

本文选题：膀胱肿瘤 + 基因沉默　；参考：《肿瘤》2016年08期

【摘要】：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(P0.05)。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(P0.05),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。
[Abstract]:Aim: to detect the expression of 1(astrocyte elevated gene-1 gene AEG-1 in human bladder cancer cells, and to investigate the effect of down-regulation of AEG-1 expression on the proliferation and invasion of T24 cells. Methods: Western blot and immunofluorescence staining were used to detect the expression of AEG-1 protein in normal bladder urinary tract epithelial cells (SV-HUC-1) and bladder cancer cell lines RT4, J82, 5637 and T24. Human bladder cancer T24 cells with high expression of AEG-1 were infected with sh RNA recombinant virus targeting AEG-1 gene specifically. Stable infected cell lines were obtained by purine mycin screening. The silencing effect of AEG-1 gene was verified by Western blotting. Then, CCK-8 assay was used. The effects of AEG-1 gene silencing on the proliferation, migration and invasion of bladder cancer T24 cells were detected by scratch healing test and Transwell chamber method. Results the expression of AEG-1 protein in the four bladder cancer cells was significantly higher than that in the normal urinary tract epithelial cells. Both 0.05).AEG-1 sh RNA recombinant virus infection could significantly down-regulate the expression of AEG-1 protein in T24 cells. Compared with T24 cells infected with sh RNA recombinant virus, the proliferation of T24 cells was significantly inhibited after down-regulation of AEG-1 expression by AEG-1 sh RNA, and the migration and invasion ability of T24 cells decreased significantly (P 0.05). Conclusion: silencing the expression of AEG-1 gene can inhibit the proliferation, migration and invasion of bladder cancer cells, suggesting that AEG-1 gene may promote the malignant biological behavior of bladder cancer cells.
【作者单位】：上海交通大学医学院附属仁济医院泌尿科;
【基金】：上海市科学技术委员会科研计划项目(编号:12ZR1417700) 上海市卫生局科研课题计划项目(编号:20134387) 上海申康医院发展中心市级医院新兴前沿技术项目(编号:SHDC12015125)
【分类号】：R737.14

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