title: Artesunate induces apoptosis of bladder cancer cells by miR-16 regulation of COX-2 expression.
abstract: Bladder cancer is the most common malignant tumor of the urinary tract and remains one of the major causes of cancer death worldwide. In this study, we investigated the effect and mechanism of Artesunate (ART), a traditional Chinese medicine, on inducing apoptosis of human bladder cancer cells. In vivo antitumor activity was investigated in bladder cancer in rat by subcutaneous injection of different concentration of ART. The effect of ART on growth inhibition and apoptosis of bladder cancer cells was evaluated using dimethylthiazoly-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis, respectively. Cyclooxygenase-2 (COX-2) and miR-16 expression levels were determined with real-time PCR. The concentrations of prostaglandin E2 (PGE2) in the supernatants of bladder cancer cells were measured with an ELISA kit. The miR-16 inhibitor or mimic were transfected into cells to up- or down-regulate miR-16 expression. ART efficiently inhibited orthotopic tumor growth in the bladder cancer rat, which is accompanied with an increase of miR-16 expression and a decrease of COX-2 expression. In vitro, ART could induce cytotoxicity and apoptosis in bladder cancer cells, but presented a much lighter toxicity effect against normal human urothelial cells. ART significantly increased miR-16 expression and decreased the expression of COX-2 and the production of PGE2. More importantly, down-regulation of miR-16 expression could reverse the effect of ART on apoptosis and COX-2 expression in bladder cells. Moreover, exogenous PGE2 could inhibit apoptosis of bladder cancer cells treated with ART. In conclusion, ART can elicit an anti-tumor effect against bladder cancer by up-regulation of miR-16 expression, which resulted in the decrease of COX-2 expression and PGE2 production. Hence, ART might be an effective drug for the treatment of bladder cancer.
authors.affiliation: Department of Urinary Surgery, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China. zuoweijp@yahoo.co.jp.
authors.forename: Wei
authors.initials: W
authors.lastname: Zuo
authors.affiliation: Department of Urinary Surgery, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China. zhenzhongwang001@gmail.com.
authors.forename: Zhen-Zhong
authors.initials: ZZ
authors.lastname: Wang
authors.affiliation: Department of Urinary Surgery, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China. junxue456@gmail.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Xue
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D037621:Artemisinins / Q000494:pharmacology* / Q000627:therapeutic use*; D000077332:Artesunate; D002460:Cell Line; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism*
chemical_list: D037621:Artemisinins; D035683:MicroRNAs; D000077332:Artesunate; D051546:Cyclooxygenase 2