Lentivirus-mediated RNA interference of clusterin enhances the chemosensitivity of EJ bladder cancer cells to epirubicin in vitro
Lentivirus-mediated RNA interference of clusterin enhances the chemosensitivity of EJ bladder cancer cells to epirubicin in vitro
复制标题
慢病毒介导的簇蛋白RNA干扰增强EJ膀胱癌细胞对表柔比星的体外化疗敏感性
DOI：
10.3892/mmr.2012.1017
复制
发表时间：
2012-11-01
影响因子：
3.4
通讯作者：
Xie, Dan
中科院分区：
文献类型：
--
作者：
Lu, Jian;Luo, Jun-Hang;Xie, Dan
异丙肾上腺素（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化学抗性密切相关。然而，CLU表达对膀胱癌对表阿霉素耐药性的影响仍然未知。本研究旨在阐明CLU在膀胱癌细胞对表阿霉素耐药中的作用。应用慢病毒介导的RNA干扰来敲低EJ膀胱癌细胞中的CLU。通过RT-PCR和western blot分析检测其效率。在稳定的CLU沉默后，建立EJ细胞系，并用或不用表阿霉素处理细胞。分别采用MTT法、创伤愈合实验、Matrigel侵袭实验、平板克隆形成实验和流式细胞术检测细胞活力、迁移能力、侵袭能力、克隆形成和细胞周期进程。结果表明，慢病毒介导的RNA干扰有效地沉默CLU在RNA和蛋白质水平。CLU敲低增加表阿霉素对EJ膀胱癌细胞的细胞毒性。慢病毒介导的靶向CLU和表阿霉素的shRNA联合治疗对膀胱癌细胞的细胞活力、迁移、侵袭力和克隆形成能力具有最大的影响。此外，细胞周期分析表明，CLU敲低增强了表阿霉素对G 0/G1细胞周期阻滞的功效。总之，我们的结果表明，CLU沉默增强EJ膀胱癌细胞对表阿霉素的化学敏感性。慢病毒介导的靶向CLU的shRNA可能是治疗膀胱癌的另一种方法。
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.