The Role of c-FLIP in Cisplatin Resistance of Human Bladder Cancer Cells
The Role of c-FLIP in Cisplatin Resistance of Human Bladder Cancer Cells
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DOI：
10.1016/j.juro.2013.01.003
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发表时间：
2013-06-01
影响因子：
6.6
通讯作者：
Lee, Sang Eun
中科院分区：
文献类型：
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作者：
Lee, Sangchul;Yoon, Cheol Yong;Lee, Sang Eun
目的：我们研究了人膀胱癌细胞顺铂耐药的机制，为顺铂耐药的膀胱癌的治疗提供新的分子靶点。材料和方法：顺铂敏感（T24）和耐药（T24 R2）的人膀胱癌细胞系的差异基因表达进行了分析和验证芯片和蛋白质印迹分析。使用细胞计数试剂盒-8测定法（Dojindo Molecular Technologies，盖瑟斯堡，马里兰州）和蛋白质印迹法评估T24 R2细胞中c-FLIP敲低引起的顺铂敏感性变化和相关机制。结果：基因芯片分析显示，在T24 R2和T24细胞系中，分别有1,086和322个基因的表达发生了2倍和4倍以上的变化。尤其是与c-FLIP相关的死亡受体凋亡途径相关的基因，包括caspase 2和9、NF-κ B、BID、c-FLIP、XIAP和cIAP 1和2，在2种细胞系中显示出差异表达。Western blot显示顺铂完全抑制T24细胞中c-FLIP的表达，但在相同剂量范围的顺铂处理后，在T24 R2细胞中未观察到c-FLIP表达的变化。通过siRNA转染抑制T24 R2细胞中c-FLIP表达，使这些细胞对顺铂治疗的敏感性显著高于未转染的T24 R2细胞（p
Purpose: We investigated the mechanisms underlying cisplatin resistance in human bladder cancer cells to provide novel molecular targets for the treatment of cisplatin resistant bladder cancer.Materials and Methods: The differential gene expression of cisplatin sensitive (T24) and resistant (T24R2) human bladder cancer cell lines was analyzed and validated by microarray and Western blot analysis. Changes in cisplatin sensitivity by c-FLIP knockdown and related mechanisms in T24R2 cells were assessed using the Cell Counting Kit-8 assay (Dojindo Molecular Technologies, Gaithersburg, Maryland) and Western blot. siRNA oligonucleotides that specifically target c-FLIP were prepared and siRNA transfection was done.Results: Microarray analysis revealed that the expression of 1,086 and 322 genes showed more than twofold and fourfold changes in the T24R2 and T24 cell lines, respectively. Especially genes involved in the c-FLIP related death receptor apoptosis pathway, including caspase 2 and 9, NF-kB, BID, c-FLIP, XIAP, and cIAP1 and 2, showed differential expression in the 2 cell lines. Western blot demonstrated complete cisplatin mediated suppression of c-FLIP expression in T24 cells but no change in c-FLIP expression was observed in T24R2 cells after cisplatin treatment in the same dose range. Suppression of c-FLIP expression in T24R2 cells by siRNA transfection rendered these cells significantly more sensitive to cisplatin treatment than untransfected T24R2 cells (p