Altered beta-tubulin isotype expression in paclitaxel-resistant human prostate carcinoma cells.
Altered beta-tubulin isotype expression in paclitaxel-resistant human prostate carcinoma cells.
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DOI：
10.1038/bjc.1998.91
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发表时间：
1998-02
影响因子：
8.8
通讯作者：
Hudes, GR
中科院分区：
文献类型：
--
作者：
Ranganathan, S;Benetatos, CA;Colarusso, PJ;Dexter, DW;Hudes, GR
为了研究β -微管蛋白同型组成在抗微管药物紫杉醇耐药性中的作用，我们将人前列腺癌（DU-145）细胞间歇性暴露于浓度增加的紫杉醇中。选择并维持在10 nM紫杉醇（Pac-10）下的细胞对该药产生了5倍的耐药。Pac-10细胞积累放射性标记紫杉醇的程度与DU-145细胞相同，耐多药-1阴性。流式细胞术分析Pac-10和DU-145细胞的细胞周期模式相似。免疫荧光染色显示，与DU-145细胞相比，Pac-10细胞中α -和β -微管蛋白水平总体升高。通过免疫荧光和western blot分析，对β -微管蛋白同型组成的检测显示，耐药细胞中β -微管蛋白同型显著增加。逆转录聚合酶链反应（RT-PCR）分析同种型证实了betaIII的增加，betaIII mRNA水平增加了9倍，betaIVa转录物也增加了5倍。此外，对增加药物水平（pac2、4、6、8和10）的紫杉醇耐药细胞的分析显示，β ii水平的增加与紫杉醇耐药性的增加呈正相关。特异性β -微管蛋白同型表达的增加和随后的微管掺入可能会改变细胞微管动力学，为紫杉醇和其他微管蛋白结合药物的抗微管作用提供防御。
To investigate the role of beta-tubulin isotype composition in resistance to paclitaxel, an anti-microtubule agent, human prostate carcinoma (DU-145) cells were intermittently exposed to increasing concentrations of paclitaxel. Cells that were selected and maintained at 10 nM paclitaxel (Pac-10) were fivefold resistant to the drug. Pac-10 cells accumulated radiolabelled paclitaxel to the same extent as DU-145 cells and were negative for MDR-1. Analysis of Pac-10 and DU-145 cells by flow cytometry showed similar cell cycle patterns. Immunofluorescent staining revealed an overall increase of alpha- and beta-tubulin levels in Pac-10 cells compared with DU-145 cells. Examination of beta-tubulin isotype composition revealed a significant increase in betaIII isotype in the resistant cells, both by immunofluorescence and by western blot analysis. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of the isotypes confirmed the increase observed for the betaIII by exhibiting ninefold higher betaIII mRNA levels and also showed fivefold increase of the betaIVa transcript. In addition, analysis of paclitaxel-resistant cells that were selected at increasing levels of the drug (Pac 2, 4, 6, 8 and 10) exhibited a positive correlation between increasing betaIII levels and increasing resistance to paclitaxel. Increased expression of specific beta-tubulin isotypes and subsequent incorporation into microtubules may alter cellular microtubule dynamics, providing a defence against the anti-microtubule effects of paclitaxel and other tubulin-binding drugs.
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DOI：
10.1073/pnas.91.24.11358
发表时间：
1994-11-22
影响因子：
11.1
作者：
PANDA, D;MILLER, HP;WILSON, L
通讯作者：
WILSON, L
影响因子：
2.9
作者：
Derry, WB;Wilson, L;Jordan, MA
通讯作者：
Jordan, MA
影响因子：
2.9
作者：
CHOMCZYNSKI, P;SACCHI, N
通讯作者：
SACCHI, N
DOI：
10.1073/pnas.90.20.9552
发表时间：
1993-10-15
影响因子：
11.1
作者：
JORDAN, MA;TOSO, RJ;WILSON, L
通讯作者：
WILSON, L
影响因子：
3.3
作者：
DHAMODHARAN, R;JORDAN, MA;WADSWORTH, P
通讯作者：
WADSWORTH, P