Bufalin induces G₀/G₁ phase arrest through inhibiting the levels of cyclin D, cyclin E, CDK2 and CDK4, and triggers apoptosis via mitochondrial signaling pathway in T24 human bladder cancer cells

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• 作者：Wen-Wen Huang^a ; ¹ ; Jai-Sing Yang^b ; ¹ ; Shu-Jen Pai^a ; Ping-Ping Wu^c ; Shu-Jen Chang^c ; Fu-Shin Chueh^d ; Ming-Jen Fan^e ; Shang-Ming Chiou^f ; ^g ; Hsiu-Maan Kuo^h ; Chin-Chung Yehⁱ ; Po-Yuan Chen^a ; Minoru Tsuzuki^j ; ^k ; Jing-Gung Chung^a ; ^e ; ^{jgchung@mail.cmu.edu.tw}
• 关键词：AIF ; apoptosis-inducing factor ; CDK ; cyclin-dependent kinase ; CsA ; cyclosporine A ; DCFH-DA ; 2&prime ; -7&prime ; -dichlorfluorescein-diacetate ; DiOC6 ; 3 ; 3&prime ; -dihexyloxacarbocyanine iodide ; DMSO ; dimethyl sulfoxide ; ECL ; enzyme chemiluminescence ; FCS ; feta
• 刊名：Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis
• 出版年：2012
• 期刊代码：115_00275107
• 类别：bio
• 出版时间：1 April, 2012
• 卷：732
• 期：1-2
• 页码：26-33
• 文件大小：1162 K

摘要

Most of the chemotherapy treatments for bladder cancer aim to kill the cancer cells, but a high recurrence rate after medical treatments is still occurred. Bufalin from the skin and parotid venom glands of toad has been shown to induce apoptotic cell death in many types of cancer cell lines. However, there is no report addressing that bufalin induced cell death in human bladder cancer cells. The purpose of this study was investigated the mechanisms of bufalin-induced apoptosis in a human bladder cancer cell line (T24). We demonstrated the effects of bufalin on the cell growth and apoptosis in T24 cells by using DAPI/TUNEL double staining, a PI exclusion and flow cytometric analysis. The effects of bufalin on the production of reactive oxygen species (ROS), the level of mitochondrial membrane potential (螖唯_m), and DNA content including sub-G1 (apoptosis) in T24 cells were also determined by flow cytometry. Western blot analysis was used to examine the expression of G₀/G₁ phase-regulated and apoptosis-associated protein levels in bufalin-treated T24 cells. The results indicated that bufalin significantly decreased the percentage of viability, induced the G₀/G₁ phase arrest and triggered apoptosis in T24 cells. The down-regulation of the protein levels for cyclin D, CDK4, cyclin E, CDK2, phospho-Rb, phospho-AKT and Bcl-2 with the simultaneous up-regulation of the cytochrome c, Apaf-1, AIF, caspase-3, -7 and -9 and Bax protein expressions and caspase activities were observed in T24 cells after bufalin treatment. Based on our results, bufalin induces apoptotic cell death in T24 cells through suppressing AKT activity and anti-apoptotic Bcl-2 protein as well as inducing pro-apoptotic Bax protein. The levels of caspase-3, -7 and -9 are also mediated apoptosis in bufalin-treated T24 cells. Therefore, bufalin might be used as a therapeutic agent for the treatment of human bladder cancer in the future.