Acquisition of docetaxel resistance in breast cancer cells reveals upregulation of ABCB1 expression as a key mediator of resistance accompanied by discrete upregulation of other specific genes and pathways
Acquisition of docetaxel resistance in breast cancer cells reveals upregulation of ABCB1 expression as a key mediator of resistance accompanied by discrete upregulation of other specific genes and pathways
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DOI：
10.1007/s13277-015-3072-4
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发表时间：
2015-06-01
期刊：
影响因子：
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通讯作者：
Schrohl, Anne-Sofie
中科院分区：
文献类型：
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作者：
Hansen, Stine Ninel;Westergaard, David;Schrohl, Anne-Sofie
微管靶向紫杉烷类药物在乳腺癌治疗中很重要，但尚未确定具有足够科学证据的预测性生物标志物以允许临床常规使用。本研究的目的是开发一个基于细胞培养的多西紫杉醇耐药发现平台，从而确定多西紫杉醇耐药的关键分子机制和预测分子特征。两种多西紫杉醇抗性细胞系 MCF7(RES) 和 MDA(RES) 是通过在 15 个月内以多西紫杉醇剂量增量逐步选择，从其各自的亲本细胞系 MCF-7 和 MDA-MB-231 产生的。细胞系对多西紫杉醇和其他化疗药物的敏感性进行了表征，并进行了全转录组 mRNA 微阵列分析。 MCF7(RES)和MDA(RES)在多西紫杉醇浓度增加时表现出双相生长抑制模式。基因表达分析指出，编码通透性糖蛋白 (Pgp) 的 ABCB1 是 MCF7(RES) 和 MDA(RES) 中最上调的基因。功能验证表明，Pgp 在低多西紫杉醇浓度（第一阶段反应）下是一个关键的耐药介质，而在较高多西紫杉醇浓度（第二阶段反应）下，其他耐药机制似乎很突出。基因表达谱表明了其他耐药机制，包括 MCF7(RES) 中的干扰素诱导蛋白家族中的基因和 MDA(RES) 中的癌睾丸抗原家族中的基因。此外，在两种耐药细胞系中，各种 ABC 转运蛋白、ECM 相关蛋白和溶酶体蛋白的表达上调。最后，MCF7(RES)和MDA(RES)对表阿霉素表现出交叉耐药性，但只有MDA(RES)对奥沙利铂表现出交叉耐药性。总之，Pgp 被确定为对低多西紫杉醇浓度耐药的关键介质，而其他耐药机制在较高多西紫杉醇浓度下也很突出。 MCF7(RES) 和 MDA(RES) 都对表阿霉素（Pgp 底物）具有交叉耐药性，但只有 MDA(RES) 对奥沙利铂（非 Pgp 底物）具有交叉耐药性，这支持 Pgp 上调是紫杉烷耐药的主要机制，而细胞系特异性改变是另一种主要机制。
The microtubule-targeting taxanes are important in breast cancer therapy, but no predictive biomarkers have yet been identified with sufficient scientific evidence to allow clinical routine use. The purposes of the present study were to develop a cell-culture-based discovery platform for docetaxel resistance and thereby identify key molecular mechanisms and predictive molecular characteristics to docetaxel resistance. Two docetaxel-resistant cell lines, MCF7(RES) and MDA(RES), were generated from their respective parental cell lines MCF-7 and MDA-MB-231 by stepwise selection in docetaxel dose increments over 15 months. The cell lines were characterized regarding sensitivity to docetaxel and other chemotherapeutics and subjected to transcriptome-wide mRNA microarray profiling. MCF7(RES) and MDA(RES) exhibited a biphasic growth inhibition pattern at increasing docetaxel concentrations. Gene expression analysis singled out ABCB1, which encodes permeability glycoprotein (Pgp), as the top upregulated gene in both MCF7(RES) and MDA(RES). Functional validation revealed Pgp as a key resistance mediator at low docetaxel concentrations (first-phase response), whereas additional resistance mechanisms appeared to be prominent at higher docetaxel concentrations (second-phase response). Additional resistance mechanisms were indicated by gene expression profiling, including genes in the interferon-inducible protein family in MCF7(RES) and cancer testis antigen family in MDA(RES). Also, upregulated expression of various ABC transporters, ECM-associated proteins, and lysosomal proteins was identified in both resistant cell lines. Finally, MCF7(RES) and MDA(RES) presented with cross-resistance to epirubicin, but only MDA(RES) showed cross-resistance to oxaliplatin. In conclusion, Pgp was identified as a key mediator of resistance to low docetaxel concentrations with other resistance mechanisms prominent at higher docetaxel concentrations. Supporting Pgp upregulation as one major mechanism of taxane resistance and cell-line-specific alterations as another, both MCF7(RES) and MDA(RES) were cross-resistant to epirubicin (Pgp substrate), but only MDA(RES) was cross-resistant to oxaliplatin (non-Pgp substrate).