miR-630 targets IGF1R to regulate response to HER-targeting drugs and overall cancer cell progression in HER2 over-expressing breast cancer.
miR-630 targets IGF1R to regulate response to HER-targeting drugs and overall cancer cell progression in HER2 over-expressing breast cancer.
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DOI：
10.1186/1476-4598-13-71
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发表时间：
2014-03-24
期刊：
影响因子：
37.3
通讯作者：
O'Driscoll L
中科院分区：
文献类型：
--
作者：
Corcoran C;Rani S;Breslin S;Gogarty M;Ghobrial IM;Crown J;O'Driscoll L
虽然用最近的HER靶向药物治疗HER 2过表达乳腺癌对一些患者非常有效，但原发性（也称为先天性）或获得性耐药限制了这些药物的成功。microRNA具有作为诊断、预后和预测生物标志物以及替代疗法的潜力。在这里，我们研究了microRNA-630（miR-630）在乳腺癌进展中的作用，并作为对HER靶向药物反应的预测生物标志物，最终产生作为治疗方法的潜力，为这些药物增加价值。我们使用qPCR研究了对HER靶向拉帕替尼和来那替尼具有先天性或获得性耐药性的乳腺癌细胞系的细胞和条件培养基中细胞内和细胞外miR-630的水平，并与相应的药物敏感细胞系进行了比较。为了支持miR-630在乳腺癌中的作用，我们研究了这种miRNA在乳腺癌肿瘤与匹配的癌旁组织中的临床相关性。使用miR-630模拟物和抑制剂的转染来操纵miR-630的表达，以评估对HER靶向药物（拉帕替尼、来那替尼和阿法替尼）应答的影响。其他与细胞侵袭性相关的表型变化通过运动性、侵袭性和失巢凋亡测定进行评价。使用TargetScan预测软件、qPCR、免疫印迹和ELISA来评估miR-630对mRNA、蛋白质及其磷酸化形式的调节。我们确定，将miR-630引入对HER-药物具有先天性或获得性耐药的细胞中可显著恢复拉帕替尼、来那替尼和阿法替尼的疗效;通过我们确定的至少部分涉及miR-630对IGF 1 R的调节的机制。相反，我们证明阻断miR-630诱导了对这些药物的耐药性/不敏感性。细胞运动性、侵袭性和失巢凋亡也被观察到通过miR-630操作显著改变，由此将miR-630引入细胞中降低了细胞侵袭性，而抑制miR-630诱导了更具侵袭性的细胞表型。综上所述，我们的研究结果表明，miR-630是HER 2过表达乳腺癌中癌细胞进展的关键调节因子，通过靶向IGF 1 R。这项研究支持miR-630作为对HER靶向药物反应的诊断和预测生物标志物，并表明治疗性添加miR-630可增强和改善患者对HER靶向药物的反应。
While the treatment of HER2 over-expressing breast cancer with recent HER-targeted drugs has been highly effective for some patients, primary (also known as innate) or acquired resistance limits the success of these drugs. microRNAs have potential as diagnostic, prognostic and predictive biomarkers, as well as replacement therapies. Here we investigated the role of microRNA-630 (miR-630) in breast cancer progression and as a predictive biomarker for response to HER-targeting drugs, ultimately yielding potential as a therapeutic approach to add value to these drugs. We investigated the levels of intra- and extracellular miR-630 in cells and conditioned media from breast cancer cell lines with either innate- or acquired- resistance to HER-targeting lapatinib and neratinib, compared to their corresponding drug sensitive cell lines, using qPCR. To support the role of miR-630 in breast cancer, we examined the clinical relevance of this miRNA in breast cancer tumours versus matched peritumours. Transfection of miR-630 mimics and inhibitors was used to manipulate the expression of miR-630 to assess effects on response to HER-targeting drugs (lapatinib, neratinib and afatinib). Other phenotypic changes associated with cellular aggressiveness were evaluated by motility, invasion and anoikis assays. TargetScan prediction software, qPCR, immunoblotting and ELISAs, were used to assess miR-630’s regulation of mRNA, proteins and their phosphorylated forms. We established that introducing miR-630 into cells with innate- or acquired- resistance to HER-drugs significantly restored the efficacy of lapatinib, neratinib and afatinib; through a mechanism which we have determined to, at least partly, involve miR-630’s regulation of IGF1R. Conversely, we demonstrated that blocking miR-630 induced resistance/insensitivity to these drugs. Cellular motility, invasion, and anoikis were also observed as significantly altered by miR-630 manipulation, whereby introducing miR-630 into cells reduced cellular aggression while inhibition of miR-630 induced a more aggressive cellular phenotype. Taken together, our findings suggest miR-630 as a key regulator of cancer cell progression in HER2 over-expressing breast cancer, through targeting of IGF1R. This study supports miR-630 as a diagnostic and a predictive biomarker for response to HER-targeted drugs and indicates that the therapeutic addition of miR-630 may enhance and improve patients’ response to HER-targeting drugs.