IN-VITRO MODULATION OF CISPLATIN ACCUMULATION IN HUMAN OVARIAN-CARCINOMA CELLS BY PHARMACOLOGICAL ALTERATION OF MICROTUBULES
IN-VITRO MODULATION OF CISPLATIN ACCUMULATION IN HUMAN OVARIAN-CARCINOMA CELLS BY PHARMACOLOGICAL ALTERATION OF MICROTUBULES
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DOI：
10.1172/jci116585
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发表时间：
1993-07-01
影响因子：
15.9
通讯作者：
HOWELL, SB
中科院分区：
文献类型：
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作者：
CHRISTEN, RD;JEKUNEN, AP;HOWELL, SB
我们以前已经表明，毛喉素和3-异丁基-1-甲基黄嘌呤（IBMX）增加顺铂（DDP）在DDP敏感的2008年人卵巢癌细胞中的积累，与其增加cAMP的能力成比例。由于cAMP的主要功能是激活蛋白激酶A，因此证实DDP蓄积的刺激是由蛋白激酶A底物介导的。我们现在表明，2008细胞暴露于毛喉素导致一个突出的52 kD膜蛋白磷酸化。对条带的微测序证明其是人β-微管蛋白。同样，用微管稳定药物紫杉醇预处理2008细胞以剂量依赖性方式增加铂积累。在11倍DDP耐药的2008/C13*5.25细胞中，相对于2008细胞，DDP蓄积减少与微管束自发形成增强以及β-微管蛋白和微管蛋白相关p53抗癌基因表达降低相关。2008/C13*5.25细胞对微管蛋白结合药物的敏感性改变，对紫杉醇过敏，对秋水仙碱交叉耐药。我们得出结论，微管蛋白的药理学改变增强了DDP的蓄积，并且2008/C13*5.25细胞中的DDP耐药表型与微管蛋白异常相关。
We have previously shown that forskolin and 3-isobutyl-1-methylxanthine (IBMX) increased accumulation of cisplatin (DDP) in DDP-sensitive 2008 human ovarian carcinoma cells in proportion to their ability to increase cAMP. Since the major function of cAMP is to activate protein kinase A, it was conjectured that the stimulation of DDP accumulation was mediated by a protein kinase A substrate. We now show that exposure of 2008 cells to forskolin resulted in phosphorylation of a prominent 52-kD membrane protein. Microsequencing of the band demonstrated it to be human beta-tubulin. Similarly, pretreatment of 2008 cells with the microtubule stabilizing drug taxol increased platinum accumulation in a dose-dependent manner. In 11-fold DDP-resistant 2008/C13*5.25 cells, decreased DDP accumulation was associated with enhanced spontaneous formation of microtubule bundles and decreased expression of beta-tubulin and the tubulin-associated p53 antioncogene relative to 2008 cells. 2008/C13*5.25 cells had altered sensitivity to tubulin-binding drugs, being hypersensitive to taxol and cross-resistant to colchicine. We conclude that pharmacologic alterations of tubulin enhance accumulation of DDP, and that the DDP-resistant phenotype in 2008/C13*5.25 cells is associated with tubulin abnormalities.