Peloruside- and laulimalide-resistant human ovarian carcinoma cells have βI-tubulin mutations and altered expression of βII- and βIII-tubulin isotypes.
Peloruside- and laulimalide-resistant human ovarian carcinoma cells have βI-tubulin mutations and altered expression of βII- and βIII-tubulin isotypes.
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DOI：
10.1158/1535-7163.mct-10-1057
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发表时间：
2011-08
影响因子：
5.7
通讯作者：
Miller JH
中科院分区：
文献类型：
--
作者：
Kanakkanthara A;Wilmes A;O'Brate A;Escuin D;Chan A;Gjyrezi A;Crawford J;Rawson P;Kivell B;Northcote PT;Hamel E;Giannakakou P;Miller JH
Peloruside A 和 laulimalide 是有效的微管稳定天然产物，其作用机制与紫杉醇相似。然而，peloruside A 和月桂马利特在微管蛋白上的结合位点仍然知之甚少。抗癌治疗中的耐药性是一个严重的问题。我们通过选择能够在其中一种药物存在下生长的 1A9 人卵巢癌细胞，开发了 Peloruside A 和月桂马内酯耐药细胞系。 1A9-laulimalide 抗性细胞 (L4) 对选择剂具有 39 倍抗性，对 peloruside A 具有 39 倍交叉抗性，而 1A9-peloruside A 抗性细胞 (R1) 对选择剂具有 6 倍抗性，同时仍对月桂马内酯敏感。两种细胞系均未表现出对紫杉醇或其他与 β-微管蛋白上的紫杉烷位点结合的药物的耐药性，也未表现出对微管不稳定药物的耐药性。耐药细胞表现出 Peloruside A/laulimalide 诱导的微管蛋白聚合受损和有丝分裂停滞受损。在 βI-微管蛋白同种型、L4 细胞的 R306H 或 R306C 以及 R1 细胞的 A296T 中发现微管蛋白突变。这是第一个基于细胞的证据支持 pelosuside A 和 laulimalide 的 β-微管蛋白结合位点。为了确定细胞的不同耐药表型是否可归因于任何其他微管蛋白改变，检查了细胞的β-微管蛋白同种型组成。仅在 L4 细胞中观察到 βII-和 βIII-微管蛋白表达增加。这些结果提供了关于微管蛋白的改变如何导致两种药物（peloruside A 和 laulimalide）独特的耐药性的见解，这两种药物具有相似的作用模式。
Peloruside A and laulimalide are potent microtubule-stabilizing natural products with a mechanism of action similar to that of paclitaxel. However, the binding site of peloruside A and laulimalide on tubulin remains poorly understood. Drug resistance in anticancer treatment is a serious problem. We developed peloruside A- and laulimalide-resistant cell lines by selecting 1A9 human ovarian carcinoma cells that were able to grow in the presence of one of these agents. The 1A9-laulimalide resistant cells (L4) were 39-fold resistant to the selecting agent and 39-fold cross-resistant to peloruside A, whereas the 1A9-peloruside A resistant cells (R1) were 6-fold resistant to the selecting agent while they remained sensitive to laulimalide. Neither cell line showed resistance to paclitaxel or other drugs that bind to the taxoid site on β-tubulin nor was there resistance to microtubule-destabilizing drugs. The resistant cells exhibited impaired peloruside A/laulimalide-induced tubulin polymerization and impaired mitotic arrest. Tubulin mutations were found in the βI-tubulin isotype, R306H or R306C for L4 and A296T for R1 cells. This is the first cell-based evidence to support a β-tubulin–binding site for peloruside A and laulimalide. To determine whether the different resistance phenotypes of the cells were attributable to any other tubulin alterations, the β-tubulin isotype composition of the cells was examined. Increased expression of βII- and βIII-tubulin was observed in L4 cells only. These results provide insight into how alterations in tubulin lead to unique resistance profiles for two drugs, peloruside A and laulimalide, that have a similar mode of action.