Clusterin、XIAP和survivin在喜树碱（CPT）耐药的PC-3细胞和CPT敏感的LNCaP细胞中的表达及其经CPT处理后的变化。

Expression of clusterin, XIAP and survivin, and their changes by camptothecin (CPT) treatment in CPT-resistant PC-3 and CPT-sensitive LNCaP cells.

作者信息

Mizutani K, Matsumoto K, Hasegawa N, Deguchi T, Nozawa Y

机构信息

Department of Urology, Gifu University Graduate school of Medicine, 1-1 Yanagido, Gifu, Gifu 5011194, Japan.

出版信息

Exp Oncol. 2006 Sep;28(3):209-15.

PMID:17080014

Abstract

AIM

Clusterin and IAPs (inhibitor of apoptosis proteins), such as survivin and XIAP, are known to be related to chemo-resistance in several cancer cells. In the current study, we investigated their expression levels in human prostate cancer cell lines, LNCaP and PC-3 which are sensitive and resistant to camptothecin (CPT), topoisomerase I inhibitor, respectively.

METHODS

LNCaP and PC-3 cells were cultured in the presence of CPT, cell death was evaluated using Hoechst 33342 and propidium iodide (PI) double staining. The expression of clusterin, XIAP and survivin on mRNA and protein levels was investigated by semi-quantitative RT-PCR and Western blotting, respectively.

RESULTS

Our data showed that 24 h treatment of LNCaP cells with 0.5 and 3.0 microM CPT resulted in higher number of apoptotic cells, than that in PC-3 cells. Western blot analysis revealed that the clusterin level in PC-3 cells was 5-fold higher than that in LNCaP cells. In contrast, XIAP expression level in PC-3 cells was lower than that in LNCaP cells, and survivin levels were similar in these two cell lines. Treatment with 0.5 and 3.0 microM CPT resulted in the reduced survivin and XIAP expression in both cell lines, while clusterin expression remained unchanged in LNCaP cells, but was increased in PC-3 cells.

CONCLUSION

The results suggest that clusterin may take a greater part in CPT-resistance than survivin and XIAP in PC-3 cells.

摘要

目的

已知聚集素和凋亡抑制蛋白（IAPs），如生存素和X连锁凋亡抑制蛋白（XIAP），与多种癌细胞的化疗耐药性相关。在本研究中，我们调查了它们在人前列腺癌细胞系LNCaP和PC-3中的表达水平，这两种细胞系分别对拓扑异构酶I抑制剂喜树碱（CPT）敏感和耐药。

方法

LNCaP和PC-3细胞在CPT存在的情况下培养，使用Hoechst 33342和碘化丙啶（PI）双重染色评估细胞死亡情况。分别通过半定量逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法研究聚集素、XIAP和生存素在mRNA和蛋白质水平的表达。

结果

我们的数据显示，用0.5和3.0微摩尔/升CPT处理LNCaP细胞24小时后，凋亡细胞数量比PC-3细胞更多。蛋白质免疫印迹分析显示，PC-3细胞中的聚集素水平比LNCaP细胞高5倍。相比之下，PC-3细胞中的XIAP表达水平低于LNCaP细胞，而这两种细胞系中的生存素水平相似。用0.5和3.0微摩尔/升CPT处理导致两种细胞系中的生存素和XIAP表达降低，而LNCaP细胞中的聚集素表达保持不变，但PC-3细胞中的聚集素表达增加。