Lentivirus-mediated RNA interference of clusterin enhances the chemosensitivity             of EJ bladder cancer cells to epirubicin in vitro.
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.
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.
Jian
J
Lu
Jun-Hang
JH
Luo
Jian
J
Pang
Jia-Zheng
JZ
Cao
Rong-Hai
RH
Wu
Zhu-Ting
ZT
Tong
Wei
W
Chen
Dan
D
Xie
D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D051152:Clusterin / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D015251:Epirubicin / Q000633:toxicity*; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D034622:RNA Interference / Q000187:drug effects*; D034741:RNA, Small Interfering / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology
D000903:Antibiotics, Antineoplastic; D051152:Clusterin; D034741:RNA, Small Interfering; D015251:Epirubicin