發(fā)布時間：2018-06-03 01:09

  本文選題：膀胱腫瘤 + 基因沉默　； 參考：《腫瘤》2016年08期

【摘要】：目的 :檢測星形細胞上調(diào)基因1(astrocyte elevated gene-1,AEG-1)在人膀胱癌細胞中的表達,探討AEG-1表達下調(diào)對膀胱癌T24細胞增殖和侵襲能力的影響。方法 :采用蛋白質(zhì)印跡法和免疫熒光染色法檢測正常膀胱尿路上皮細胞SV-HUC-1以及膀胱癌細胞株RT4、J82、5637和T24中AEG-1蛋白的表達水平。將特異性靶向AEG-1基因的sh RNA重組病毒感染自身高表達AEG-1的膀胱癌T24細胞,經(jīng)嘌呤霉素篩選獲得穩(wěn)定感染細胞系,采用蛋白質(zhì)印跡法驗證AEG-1基因被沉默的效果;然后,采用CCK-8法、劃痕愈合實驗和Transwell小室法分別檢測AEG-1基因沉默對膀胱癌T24細胞增殖、遷移和侵襲能力的影響。結果 :4種膀胱癌細胞中AEG-1蛋白表達水平明顯高于正常膀胱尿路上皮細胞(P值均0.05)。AEG-1 sh RNA重組病毒感染能明顯下調(diào)膀胱癌T24細胞中AEG-1蛋白的表達(P0.05)。與感染陰性對照sh RNA重組病毒的T24細胞相比,AEG-1 sh RNA下調(diào)AEG-1表達后T24細胞的增殖受到明顯抑制(P0.05),而且細胞的遷移和侵襲能力均明顯減弱(P值均0.05)。結論 :沉默AEG-1基因表達可抑制膀胱癌細胞的增殖、遷移及侵襲,提示AEG-1基因可能具有促進膀胱癌細胞發(fā)生惡性生物學行為的作用。
[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.
【作者單位】： 上海交通大學醫(yī)學院附屬仁濟醫(yī)院泌尿科;
【基金】：上海市科學技術委員會科研計劃項目(編號:12ZR1417700) 上海市衛(wèi)生局科研課題計劃項目(編號:20134387) 上海申康醫(yī)院發(fā)展中心市級醫(yī)院新興前沿技術項目(編號:SHDC12015125)
【分類號】：R737.14

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