MicroRNA-181a promotes docetaxel resistance in prostate cancer cells.
MicroRNA-181a promotes docetaxel resistance in prostate cancer cells.
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DOI：
10.1002/pros.23358
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发表时间：
2017-06
期刊：
影响因子：
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通讯作者：
Gao AC
中科院分区：
文献类型：
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作者：
Armstrong CM;Liu C;Lou W;Lombard AP;Evans CP;Gao AC
多西他赛是用于治疗去势抵抗性前列腺癌（CRPC）的主要药物之一。不幸的是，随着时间的推移，患者总是对多西他赛治疗产生耐药性，并且他们的疾病将继续进展。抗药性产生的机制仍不完全清楚。本研究旨在确定miRNA，特别是miR-181 a，参与CRPC多西他赛耐药。使用实时PCR测量亲本和多西他赛耐药C4-2B和DU 145细胞（TaxR和DU 145-DTXR）中的miR-181 a表达。通过分别用miR-181 a模拟物或反义物转染亲本或多西他赛抗性细胞来调节它们中的miR-181 a表达。转染后，在48小时后用或不用多西他赛测定细胞数。还确定了由miR-181 a诱导的对卡巴他赛的交叉抗性。蛋白质印迹法用于确定ABCB 1蛋白表达和罗丹明测定用于评估活性。在C4-2B TaxR和PC 3细胞中，通过蛋白质印迹法评估磷酸化p53表达，并通过ELISA测量细胞凋亡，这些细胞具有抑制或过表达的miR-181 a表达，有或没有多西他赛。与亲本细胞相比，miR-181 a在TaxR和DU 145-DTXR细胞中显著过表达。亲本细胞中miR-181 a的过表达赋予多西他赛和卡巴他赛抗性，并且TaxR细胞中miR-181 a的敲低使它们对多西他赛和卡巴他赛治疗重新敏感。未观察到miR-181 a影响ABCB 1的表达或活性，ABCB 1是一种先前被证明与多西他赛耐药性高度相关的蛋白质。TaxR细胞中miR-181 a的敲低诱导磷酸化p53表达。此外，单独的miR-181 a敲低诱导TaxR细胞凋亡，这可以通过添加DTX进一步增强。mir-181 a在前列腺癌细胞中的过表达有助于它们对多西他赛和卡巴他赛的抗性，并且抑制mir-181 a表达可以恢复治疗反应。这部分是由于p53磷酸化和凋亡的调节。
Docetaxel is one of the primary drugs used for treating castration resistant prostate cancer (CRPC). Unfortunately, over time patients invariably develop resistance to docetaxel therapy and their disease will continue to progress. The mechanisms by which resistance develops are still incompletely understood. This study seeks to determine the involvement of miRNAs, specifically miR-181a, in docetaxel resistance in CRPC. Real-time PCR was used to measure miR-181a expression in parental and docetaxel resistant C4-2B and DU145 cells (TaxR and DU145-DTXR). miR-181a expression was modulated in parental or docetaxel resistant cells by transfecting them with miR-181a mimics or antisense, respectively. Following transfection, cell number was determined after 48 h with or without docetaxel. Cross resistance to cabazitaxel induced by miR-181a was also determined. Western blots were used to determine ABCB1 protein expression and rhodamine assays used to assess activity. Phospho-p53 expression was assessed by western blot and apoptosis was measured by ELISA in C4-2B TaxR and PC3 cells with inhibited or overexpressed miR-181a expression with or without docetaxel. miR-181a is significantly overexpressed in TaxR and DU145-DTXR cells compared to parental cells. Overexpression of miR-181a in parental cells confers docetaxel and cabazitaxel resistance and knockdown of miR-181a in TaxR cells re-sensitizes them to treatment with both docetaxel and cabazitaxel. miR-181a was not observed to impact ABCB1 expression or activity, a protein which was previously demonstrated to be highly involved in docetaxel resistance. Knockdown of miR-181a in TaxR cells induced phospho-p53 expression. Furthermore, miR-181a knockdown alone induced apoptosis in TaxR cells which could be further enhanced by the addition of DTX. Overexpression of mir-181a in prostate cancer cells contributes to their resistance to docetaxel and cabazitaxel and inhibition of mir-181a expression can restore treatment response. This is due, in part, to modulation of p53 phosphorylation and apoptosis.