Chemosensitization of gemcitabine-resistant human bladder cancer cell line both in vitro and in vivo using antisense oligonucleotide targeting the anti-apoptotic gene, clusterin
Chemosensitization of gemcitabine-resistant human bladder cancer cell line both in vitro and in vivo using antisense oligonucleotide targeting the anti-apoptotic gene, clusterin
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DOI：
10.1111/j.1464-410x.2008.08098.x
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发表时间：
2009-02-01
影响因子：
4.5
通讯作者：
Gleave, Martin E.
中科院分区：
文献类型：
--
作者：
Muramaki, Mototsugu;So, Alan;Gleave, Martin E.
目的：观察持续吉西他滨治疗后膀胱癌细胞中聚簇素（scu -2）表达的变化，并确定敲低scu -2是否能重新引入吉西他滨耐药细胞对吉西他滨治疗的敏感性。将人膀胱癌细胞株UM-UC-3持续暴露于体外增加剂量的吉西他滨，培养出耐吉西他滨细胞株UM-UC-3R。然后利用针对scu -2基因的反义寡核苷酸（OGX-011）分析了scu -2在体外和体内获得的化学耐药表型中的作用。用吉西他滨处理亲代UM-UC-3细胞（UM-UC-3P）可诱导scu -2蛋白瞬间上调。与UM-UC-3P细胞相比，UM-UC-3R细胞中scu -2表达水平持续升高（6.4倍）。用OGX-011处理UM-UC-3R细胞导致了剂量依赖性和序列特异性的scu -2表达抑制。此外，OGX-011在体外使UM-UC-3R细胞对吉西他滨化学致敏，浓度降低，从100 nm到10 nm的效果降低50% （IC50）。注射UM-UC-3R细胞的裸鼠肿瘤体积和转移发生率明显大于注射UM-UC-3P细胞的裸鼠；然而，在两组中，全身给药OGX-011加低剂量吉西他滨显著抑制肿瘤体积和转移发生率。这些研究结果表明，scu -2在膀胱癌细胞化疗耐药表型的获得中起着重要作用，使用OGX-011联合化疗药物通过增强化疗敏感性来降低scu -2可能是治疗晚期膀胱癌的一种有吸引力的方法。
To characterize changes in clusterin (sCLU-2) expression in bladder cancer cells after continuous treatment with gemcitabine and to determine whether knockdown of sCLU-2 can re-introduce sensitivity of gemcitabine-resistant cells to treatment with gemcitabine.A human bladder cancer cell line, UM-UC-3, was continuously exposed to increasing doses of gemcitabine in vitro, and a gemcitabine-resistant cell line UM-UC-3R was developed. The role of sCLU-2 in chemoresistant phenotype acquired in both in vitro and in vivo was then analysed using antisense oligonucleotide targeting the sCLU-2 gene (OGX-011).Treatment of parental UM-UC-3 cells (UM-UC-3P) with gemcitabine induced transient up-regulation of sCLU-2 protein. There was a sustained increase in sCLU-2 expression levels in UM-UC-3R compared with UM-UC-3P cells (6.4-fold). Treatment of UM-UC-3R cells with OGX-011 resulted in a dose-dependent and sequence- specific inhibition in sCLU-2 expression. Furthermore, OGX-011 chemo-sensitized UM-UC-3R cells to gemcitabine in vitro with a reduction in the concentration that reduces the effect by 50% (IC50) from 100 nm to 10 nm. Tumour volume and the incidence of metastasis in nude mice injected with UM-UC-3R cells was significantly greater than those of nude mice injected with UM-UC-3P cells; however, systemic administration of OGX-011 plus a low dose of gemcitabine significantly suppressed tumour volume and the incidence of metastasis in both groups.These findings suggest that sCLU-2 plays a significant role in the acquisition of chemoresistant phenotype in bladder cancer cells and the knockdown of sCLU-2 using OGX-011 combined with a chemotherapeutic agent could be an attractive approach for advanced bladder cancer through the enhancement of chemosensitivity.