ER-α36 mediates cisplatin resistance in breast cancer cells through EGFR/HER-2/ERK signaling pathway.
ER-α36 mediates cisplatin resistance in breast cancer cells through EGFR/HER-2/ERK signaling pathway.
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ER-α 36通过EGFR/HER-2/ERK信号通路介导乳腺癌细胞顺铂耐药
DOI：
10.1186/s13046-018-0798-z
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发表时间：
2018-06-25
期刊：
影响因子：
--
通讯作者：
Chen B
中科院分区：
文献类型：
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作者：
Zhu L;Zou J;Zhao Y;Jiang X;Wang Y;Wang X;Chen B
ER-α36是一种新的ER-α66变异体，已被证明可促进乳腺癌细胞对他莫昔芬的耐药性。然而，ER-α36在乳腺癌细胞顺铂耐药中的作用和机制尚不清楚。本研究探讨ER-α36在乳腺癌细胞顺铂耐药中的表达和作用，并阐明其可能的机制。免疫印迹法检测ER-α36和非基因组雌激素信号转导相关蛋白的表达。采用CCK-8比色法、单层集落形成实验和细胞凋亡实验检测顺铂的敏感性。将ER-α36siRNAs/shRNAs和高表达载体分别导入细胞，下调或上调ER-α36的表达。通过功能丧失和功能增益分析，研究ER-α36在顺铂敏感性中的作用。应用免疫印迹法检测ER-α36与EGFR/HER-2的相互作用。建立小鼠乳腺癌移植瘤模型，验证ER-α36在体内的作用。与顺铂敏感细胞相比，ER-α36在顺铂耐药的乳腺癌细胞中的表达水平更高。顺铂以剂量依赖方式诱导乳腺癌细胞ER-α36表达上调。ER-α36过表达导致细胞对顺铂产生耐药，ER-α36在顺铂耐药的乳腺癌细胞中被敲除，恢复了顺铂的敏感性。ER-α36的上调导致非基因组雌激素信号的激活，这与顺铂耐药有关。用激酶抑制剂阻断ER-α36介导的非基因组雌激素信号显著抑制顺铂诱导的ER-α36的表达，并增加顺铂的敏感性。体内实验还证实，ER-α36的上调降低了小鼠乳腺癌异种移植模型对顺铂的敏感性。这一结果首次证实ER-α36通过非基因组雌激素信号途径介导乳腺癌细胞对顺铂的耐药，提示ER-α36可能成为顺铂耐药的新靶点和乳腺癌顺铂敏感性的潜在指标。
ER-α36, a novel ER-α66 variant, has been demonstrated to promote tamoxifen resistance in breast cancer cells. However, the role and mechanisms of ER-α36 in cisplatin resistance of breast cancer cells remain unclear. This study investigates the expression and role of ER-α36 in cisplatin resistance of breast cancer cells and elucidates its underlying mechanisms. The expression of ER-α36 and the proteins involved in nongenomic estrogen signaling was evaluated by western blot analysis. Cisplatin sensitivity was explored by CCK-8 assay, monolayer colony formation assay and apoptosis assays, respectively. ER-α36 siRNAs/shRNAs and overexpression vector were transfected into cells to down-regulate or up-regulate ER-α36 expression. Loss-and gain-of function assays were performed to investigate the role of ER-α36 in cisplatin sensitivity. The interaction between ER-α36 and EGFR/HER-2 were detected using CoIP. A mouse xenograft model of breast cancer was established to verify the role of ER-α36 in vivo. ER-α36 is expressed at higher levels in cisplatin-resistant breast cancer cells compared to cisplatin sensitive cells. Cisplatin induced up-regulation of ER-α36 in a dose-dependent manner in breast cancer cells. Overexpression of ER-α36 leaded to cell resistant to cisplatin and knockdown of ER-α36 in cisplatin-resistant breast cancer cells restored cisplatin sensitivity. The up-regulation of ER-α36 resulted in increased activation of nongenomic estrogen signaling, which was responsible for cisplatin resistance. Disruption of ER-α36-mediated nongenomic estrogen signaling with kinase inhibitors significantly inhibited cisplatin-induced expression of ER-α36 and increased cisplatin sensitivity. The in vivo experiment also confirmed that up-regulation of ER-α36 attenuated cisplatin sensitivity in a mouse xenograft model of breast cancer. The results for the first time demonstrated that ER-α36 mediates cisplatin resistance in breast cancer cells through nongenomic estrogen signaling, suggesting that ER-α36 may serve as a novel target for cisplatin resistance and a potential indicator of cisplatin sensitivity in breast cancer treatment.