Yang Fan, Fu Ziyi, Yang Mengzhu, Sun Chunxiao, Li Yongfei, Chu Jiahui, Zhang Yanhong, Li Wei, Huang Xiang, Li Jun, Wu Hao, Ding Xiaorong, Yin Yongmei
Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
The First Clinical College of Nanjing Medical University, Nanjing, China.
J Cell Physiol. 2019 Sep;234(9):16102-16113. doi: 10.1002/jcp.28268. Epub 2019 Feb 15.
Although an immense effort has been made to develop a novel biomarker for response to trastuzumab, no reliable biomarkers are available to guide management, expect for HER2. The aim of this study was to examine the relationship between microRNA (miRNA) expression and resistance to trastuzumab.
Differentially expressed miRNAs between trastuzumab-resistant and trastuzumab-sensitive cell lines were analyzed using microarrays. We performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to determine the functions of differentially expressed miRNA and their targeted genes. Furthermore, the protein-protein interactions (PPI) network was analyzed. Serum samples were collected from patients with HER2-positive breast cancer who were treated with trastuzumab. We validated the miRNAs expression levels by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) in these serums. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive performance of the miRNA.
Using miRNA microarrays, 151 miRNAs that significant differentially expressed between the trastuzumab-resistant and sensitive cells were identified, including 46 upregulated and 105 downregulated miRNAs. Results of real-time PCR confirmed seven miRNAs in cell lines. PI3K-Akt signaling pathway was involved in regulating biological function according to KEGG analysis. Compared with the serums of trastuzumab-sensitive patients, three miRNAs, namely miR-200b, miR-135b, and miR-29a, were identified to be upregulated, and miR-224 was downregulated in the trastuzumab-resistant serums. ROC analysis showed that four miRNAs were correlated with trastuzumab resistance. Furthermore, three subnetwork modules of PPI network were obtained.
The results indicated that miRNAs were reliable predictive biomarkers for response to trastuzumab.
尽管人们为开发一种用于预测曲妥珠单抗疗效的新型生物标志物付出了巨大努力，但除HER2外，尚无可靠的生物标志物可用于指导治疗。本研究旨在探讨微小RNA（miRNA）表达与曲妥珠单抗耐药性之间的关系。
使用微阵列分析曲妥珠单抗耐药和敏感细胞系之间差异表达的miRNA。我们进行了基因本体论和京都基因与基因组百科全书（KEGG）富集分析，以确定差异表达miRNA及其靶向基因的功能。此外，还分析了蛋白质-蛋白质相互作用（PPI）网络。收集接受曲妥珠单抗治疗的HER2阳性乳腺癌患者的血清样本。我们通过定量逆转录聚合酶链反应（qRT-PCR）验证了这些血清中miRNA的表达水平。进行了受试者工作特征（ROC）曲线分析，以评估miRNA的预测性能。
使用miRNA微阵列，鉴定出151个在曲妥珠单抗耐药和敏感细胞之间显著差异表达的miRNA，包括46个上调的miRNA和105个下调的miRNA。实时PCR结果在细胞系中证实了7个miRNA。根据KEGG分析，PI3K-Akt信号通路参与调节生物学功能。与曲妥珠单抗敏感患者的血清相比，在曲妥珠单抗耐药血清中鉴定出3个miRNA，即miR-200b、miR-135b和miR-29a上调，而miR-224下调。ROC分析表明，4个miRNA与曲妥珠单抗耐药相关。此外，还获得了PPI网络的3个子网络模块。
结果表明，miRNA是曲妥珠单抗疗效的可靠预测生物标志物。
