Survival of docetaxel-resistant prostate cancer cells in vitro depends on phenotype alterations and continuity of drug exposure
Survival of docetaxel-resistant prostate cancer cells in vitro depends on phenotype alterations and continuity of drug exposure
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DOI：
10.1007/s00018-002-8498-3
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发表时间：
2002-07-01
影响因子：
8
通讯作者：
Akerley, W
中科院分区：
文献类型：
--
作者：
Makarovskiy, AN;Siryaporn, E;Akerley, W
我们在体外评估了紫杉醇和多西他赛对PC-3和DU-145前列腺癌细胞系的作用，以更好地了解药物诱导的肿瘤细胞死亡的下游事件。紫杉烷处理DU-145细胞诱导快速细胞死亡的凋亡，但在PC-3细胞，治疗实现生长停滞，随后广泛的核分裂，导致多核，巨细胞形成和延迟的细胞死亡。为了确定巨大的多核细胞是否能够产生增殖和耐药的幸存者，我们首先描绘了药物活性和细胞毒性剂量范围的动力学。通过比色法和细胞活力测定对两种细胞系的分析表明连续施用紫杉烷类的细胞毒性得到改善。使用选定剂量和时间表的多西他赛诱导巨大的多核细胞，这些细胞产生对紫杉醇耐药的幸存者，这些幸存者对紫杉醇和其他化疗药物仍然敏感。回收的克隆的生长和形态与亲本细胞相似。这些克隆的免疫荧光和免疫印迹测定的耐药表型与β-微管蛋白IV亚型的瞬时表达相关，并且独立于P-糖蛋白、bcl-2和bcl-xL。耐药克隆将有助于建立紫杉烷类药物耐药进展的模型，并鉴定未知的和临床上重要的细胞死亡和耐药的分子机制。
We evaluated in vitro the effect of paclitaxel and docetaxel on PC-3 and DU-145 prostate cancer cell lines to understand better the downstream events in drug-induced tumor cell death. Taxane treatments of DU-145 cells induced rapid cell death by apoptosis, but in PC-3 cells, treatments achieved growth arrest, followed by extensive karyokinesis resulting in multinucleation, giant-cell formation and delayed cell death. To determine if the giant multinucleated cells were able to produce proliferating and drug-resistant survivors, we first delineated the kinetics of drug activity and cytotoxic dose range. Analysis of both lines by colorimetric and cell viability assays demonstrated improved cytotoxicity of taxanes applied continuously. Selected doses and schedules of docetaxel were used to induce giant multinucleated cells that gave rise to docetaxel-resistant survivors, which remained sensitive to paclitaxel and other chemotherapeutics. Growth and morphology of the recovered clones was similar to parental cells. The resistant phenotype of these clones determined by immunofluorescence and immunoblot was associated with transient expression of the beta-tubulin IV isoform and was independent of P-glycoprotein, bcl-2 and bcl-xL. Resistant clones will be useful to model progression of resistance to taxanes and to identify unknown and clinically important molecular mechanisms of cell death and resistance.