Department of Urology, The First Affiliated Hospital of Sun Yat-sen University, State Key Laboratory of Oncology in South China, Cancer Center, Sun Yat-sen University, Guangzhou 510080, PR China.
Mol Med Rep. 2012 Nov;6(5):1133-9. doi: 10.3892/mmr.2012.1017. Epub 2012 Aug 3.
Clusterin (CLU) is a glycoprotein that is over-expressed in a number of malignant tumors and has been proven to correlate closely with the chemoresistance of several cancer cells to chemotherapeutic agents. However, the effect of CLU expression on the chemoresistance of bladder cancer to epirubicin remains unknown. In the present study, we aimed to elucidate the role of CLU in the chemoresistance of bladder cancer cells to epirubicin. Lentivirus-mediated RNA interference was applied to knock down CLU in EJ bladder cancer cells. The efficiency was examined by RT-PCR and western blot analysis. After stable CLU silencing, an EJ cell line was established and cells were treated with or without epirubicin. Cell viability, migration, invasiveness, clone formation and cell cycle progression were assessed by MTT assay, wound healing assay, Matrigel invasion assay, plate clone formation assay and flow cytometry, respectively. The results indicated that lentivirus-mediated RNA interference effectively silenced CLU at the RNA and protein levels. CLU knockdown increased the cytotoxicity of epirubicin to EJ bladder cancer cells. Combined treatment with lentivirus-mediated shRNA targeting CLU and epirubicin had maximum effects in bladder cancer cells on cell viability, migration, invasiveness and clone-forming ability. Furthermore, cell cycle analysis indicated that CLU knockdown reinforced the efficacy of epirubicin on G0/G1 cell cycle arrest. Taken together, our results suggest that CLU silencing enhances chemosensitivity of EJ bladder cancer cells to epirubicin. Lentivirus-mediated shRNA targeting CLU may be an alternative approach in the treatment of bladder cancer.
簇集蛋白（CLU）是一种糖蛋白，在许多恶性肿瘤中过度表达，并且已被证明与几种癌细胞对化疗药物的耐药性密切相关。然而，CLU 表达对膀胱癌对表柔比星耐药性的影响尚不清楚。在本研究中，我们旨在阐明 CLU 在膀胱癌细胞对表柔比星耐药性中的作用。慢病毒介导的 RNA 干扰用于敲低 EJ 膀胱癌细胞中的 CLU。通过 RT-PCR 和 Western blot 分析检查效率。在稳定沉默 CLU 后，建立了 EJ 细胞系，并对细胞进行了表柔比星处理或不处理。通过 MTT 测定、划痕愈合测定、Matrigel 侵袭测定、平板克隆形成测定和流式细胞术分别评估细胞活力、迁移、侵袭、克隆形成和细胞周期进程。结果表明，慢病毒介导的 RNA 干扰可有效在 RNA 和蛋白质水平上沉默 CLU。CLU 敲低增加了表柔比星对 EJ 膀胱癌细胞的细胞毒性。用针对 CLU 的慢病毒介导 shRNA 与表柔比星联合治疗对膀胱癌细胞的细胞活力、迁移、侵袭和克隆形成能力具有最大影响。此外，细胞周期分析表明 CLU 敲低增强了表柔比星对 G0/G1 细胞周期阻滞的功效。总之，我们的结果表明，沉默 CLU 增强了 EJ 膀胱癌细胞对表柔比星的化疗敏感性。慢病毒介导的靶向 CLU 的 shRNA 可能是膀胱癌治疗的一种替代方法。
