High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth
High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth
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DOI：
10.1016/j.molonc.2013.10.001
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发表时间：
2014-02-01
影响因子：
6.6
通讯作者：
Perala, Merja
中科院分区：
文献类型：
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作者：
Leivonen, Suvi-Katri;Sahlberg, Kristine Kleivi;Perala, Merja
MicroRNAs(MiRNAs)是在转录后调节基因表达的非编码RNAs。我们已经确定了miRNAs在调节乳腺癌中人表皮生长因子受体2(HER2)途径中的作用。我们用由810个人miRNAs组成的miRNA模拟文库筛选了两个HER2扩增细胞系(KPL-4和JIMT-1)，进行了miRNA功能增益分析。用反相蛋白质芯片检测HER2、磷酸化AKT、磷酸化ERK1/2、细胞增殖(Ki67)和细胞凋亡率(CPARP)。RANK产物分析确定38个miRNAs(Q<；0.05)抑制HER2信号和细胞生长，其中最有效的是miR-491-5P、miR-634、miR-637和miR-342-5P。我们还鉴定了直接针对HER2的miRNAs，并鉴定了7个新的miRNAs(miR-552、miR-541、miR-193a-5p、miR-453、miR-134、miR-498和miR-331-3p)作为HER2 3‘非编码区的直接调控因子。我们证明了miRNAs的临床相关性，并发现在HER2阳性的乳腺肿瘤中miR-342-5p和miR-744*的表达显著下调，而来自两组乳腺癌患者的HER2阴性肿瘤(101例和1302例)中miR-342-5p和miR-744*的表达显著下调。MIR-342-5P特异性地抑制HER2阳性细胞的生长，因为它在体外对HER2阴性对照细胞的生长没有影响。此外，在两组乳腺癌患者中，miR-342-5p的高表达与更好的生存相关。总之，我们已经确定了miRNAs，它们是HER2途径的有效负调控因子，可能在体内乳腺癌进展过程中发挥作用。这些结果提供了对HER2调控的机械性见解，可能为HER2阳性乳腺癌的预防和治疗抑制开辟潜在的新策略。(C)2013年欧洲生化学会联合会。爱思唯尔出版，版权所有。
MicroRNAs (miRNAs) are non-coding RNAs regulating gene expression post-transcriptionally. We have characterized the role of miRNAs in regulating the human epidermal growth factor receptor 2 (HER2)-pathway in breast cancer. We performed miRNA gain-of-function assays by screening two HER2 amplified cell lines (KPL-4 and JIMT-1) with a miRNA mimic library consisting of 810 human miRNAs. The levels of HER2, phospho-AKT, phospho-ERK1/2, cell proliferation (Ki67) and apoptosis (cPARP) were analyzed with reverse-phase protein arrays. Rank product analyses identified 38 miRNAs (q < 0.05) as inhibitors of HER2 signaling and cell growth, the most effective being miR-491-5p, miR-634, miR-637 and miR-342-5p. We also characterized miRNAs directly targeting HER2 and identified seven novel miRNAs (miR-552, miR-541, miR-193a-5p, miR-453, miR-134, miR-498, and miR-331-3p) as direct regulators of the HER2 3'UTR. We demonstrated the clinical relevance of the miRNAs and identified miR-342-5p and miR-744* as significantly down-regulated in HER2-positive breast tumors as compared to HER2-negative tumors from two cohorts of breast cancer patients (101 and 1302 cases). miR-342-5p specifically inhibited HER2-positive cell growth, as it had no effect on the growth of HER2-negative control cells in vitro. Furthermore, higher expression of miR-342-5p was associated with better survival in both breast cancer patient cohorts. In conclusion, we have identified miRNAs which are efficient negative regulators of the HER2 pathway that may play a role in vivo during breast cancer progression. These results give mechanistic insights in HER2 regulation which may open potential new strategies towards prevention and therapeutic inhibition of HER2-positive breast cancer. (C) 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.