Stravopodis Dimitrios J, Karkoulis Panagiotis K, Konstantakou Eumorphia G, Melachroinou Sophia, Lampidonis Antonis D, Anastasiou Dimitra, Kachrilas Stefanos, Messini-Nikolaki Niki, Papassideri Issidora S, Aravantinos Gerasimos, Margaritis Lukas H, Voutsinas Gerassimos E
Department of Cell Biology and Biophysics, Faculty of Biology, University of Athens, Panepistimiopolis, Zografou, 15784 Athens, Greece.
Int J Oncol. 2009 Jan;34(1):137-60.
Doxorubicin is an important component of combination therapy for muscle-invasive urinary bladder cancer. Treatment with this topoisomerase II poison is able to interfere with cell cycle progression and lead to cancer cell death. Using FACS analysis, Western immunoblotting and semi-quantitative RT-PCR, we studied the effects of doxorubicin on cell cycle progression and apoptosis, and also explored the possibility of using groups of genes as biomarkers of prognosis and/or response to doxorubicin treatment in human urinary bladder cancer cells. Doxorubicin induced dose-dependent G2/M and/or G1/S cell cycle arrest, followed by grade- and dose-dependent reduction in the amount of the cytosolic trimeric form of FasL, activation of Caspase-8, Caspase-9, Caspase-3, cleavage of PARP, Lamin A/C, Bcl-XL/S and interestingly Hsp90, and finally cell death. Data presented here also suggest the use of the expression patterns of Cyclin-E2, Cyclin-F, p63, p73, FasL, TRAIL, Tweak, Tweak-R, XAF-1, OPG and Bok genes for identification of the differentiation grade, and Cyclin-B2, GADD45A, p73, FasL, Bik, Bim, TRAIL, Fas, Tweak-R, XAF-1, Bcl-2, Survivin, OPG, DcR2 and Bcl-XL genes for the detection of response to doxorubicin in human bladder cancer cells.
多柔比星是肌肉浸润性膀胱癌联合治疗的重要组成部分。使用这种拓扑异构酶II毒药进行治疗能够干扰细胞周期进程并导致癌细胞死亡。我们通过流式细胞术分析、蛋白质免疫印迹法和半定量逆转录-聚合酶链反应，研究了多柔比星对细胞周期进程和细胞凋亡的影响，还探讨了使用基因组合作为人类膀胱癌细胞预后和/或对多柔比星治疗反应生物标志物的可能性。多柔比星诱导剂量依赖性的G2/M和/或G1/S细胞周期阻滞，随后是FasL胞质三聚体形式数量的分级和剂量依赖性减少、半胱天冬酶-8、半胱天冬酶-9、半胱天冬酶-3的激活、聚（ADP-核糖）聚合酶、核纤层蛋白A/C、Bcl-XL/S以及有趣的热休克蛋白90的裂解，最终导致细胞死亡。此处呈现的数据还表明，可利用细胞周期蛋白-E2、细胞周期蛋白-F、p63、p73、FasL、肿瘤坏死因子相关凋亡诱导配体（TRAIL）、肿瘤坏死因子样弱凋亡诱导因子（Tweak）、Tweak受体（Tweak-R）、X连锁凋亡抑制蛋白相关因子-1（XAF-1）、骨保护素（OPG）和Bok基因的表达模式来鉴定分化程度，利用细胞周期蛋白-B2、生长停滞和DNA损伤诱导蛋白45A（GADD45A）、p73、FasL、Bik、Bim、TRAIL、Fas、Tweak-R、XAF-1、Bcl-2、生存素、OPG、诱饵受体2（DcR2）和Bcl-XL基因来检测人类膀胱癌细胞对多柔比星的反应。
