Validation and target gene screening of hsa-miR-205 in lung squamous cell carcinoma
Validation and target gene screening of hsa-miR-205 in lung squamous cell carcinoma
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DOI：
10.3760/cma.j.issn.0366-6999.20121121
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发表时间：
2014-01-20
影响因子：
6.1
通讯作者：
Lu Shaohua
中科院分区：
文献类型：
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作者：
Huang Wei;Jin Yi;Lu Shaohua
背景肺癌分为鳞状细胞癌（squamous cell carcinoma，SQ）、腺癌（adenocarcinoma，AC）和小细胞肺癌（small cell lung carcinoma，SCLC）。SQ是肺癌的主要亚型。目前，由于缺乏对其驱动癌基因的了解，没有针对SQ的靶向治疗。本研究旨在验证SQ特异性生物标志物hsa-nniR-205在中国肺癌患者中的作用，并筛选其候选靶基因，为进一步研究SQ靶向治疗提供理论依据。对197例大体解剖的（癌细胞>75%）手术肺组织（45个SQ、44个AC、54个SCLC和54个相邻正常组织）中的miR-205的表达谱以验证miR-205的表达谱。此外，通过网关miRecords预测该microRNA的靶点，并使用KEGG（京都基因和基因组百科全书）数据库将其映射到肺癌相关途径。结果MicroRNA-205对肺鳞癌和腺癌的诊断准确率较高，曲线下面积（AUC）为0.985，与正常肺组织相比，差异有显著性（P < 0.05），与正常肺组织相比，差异有显著性（P < 0.05）。以及在福尔马林固定石蜡包埋（FFPE）手术肺组织中以0.978的AUC区分SQ与SCLC。nniR-205的预测靶点与肺癌信号通路的52个关键成员相关。10个靶基因（ACSL 1、AXIN 2、CACNA 2D 2、FOXO 3、PPP 1 R3 A、PRKAG 3、RUNX 1、SMAD 4、STK 3和TBL 1XR 1）在SQ中显著下调，并与nniR-205呈强负相关，而一个靶基因（CDH 3）在SQ中表达上调，并与miR-205呈强正相关。205。此外，我们确定了11个重要的miR-205的靶基因，这些基因可用于进一步的功能研究，作为开发SQ靶向治疗的基础。
Background Lung cancers are classified as squamous cell carcinoma (SQ), adenocarcinoma (AC) and small cell lung carcinoma (SCLC). SQ is the major subtype of lung cancer. Currently, there are no targeted therapies for SQ due to lack of understanding its driving oncogenes. In this study, we validated an SQ specific biomarker hsa-nniR-205 in Chinese patients with lung cancer and screened its candidate target genes for further functional studies to enrich knowledge in SQ target therapies.Methods Quantitative reverse-transcription PCR (quantitative RT-PCR) was performed on 197 macro-dissected (cancerous cells >75%) surgical lung tissues (45 SQ, 44 AC, 54 SCLC and 54 adjacent normal tissues) to validate the expression profiles of miR-205. Furthermore, the targets of this microRNA were predicted through the gateway miRecords and mapped to lung cancer-associated pathways using the KEGG (Kyoto Encyclopedia of Genes and Genomes) database. Then quantitative RT-PCR was performed on an independent cohort of 44 snap-frozen surgical lung tissues to concurrently assess the expression profiles of miR-205 and its 52 putative targeted genes.Results MicroRNA-205 yielded high diagnostic accuracy in discriminating SQ from AC with an area under the curve (AUC) of 0.985, and discriminating SQ from SCLC with an AUC of 0.978 in formalin-fixed paraffin-embedded (FFPE) surgical lung tissues. Predicted targets of nniR-205 were associated with 52 key members of lung cancer signaling pathways. Ten target genes (ACSL1, AXIN2, CACNA2D2, FOXO3, PPP1R3A, PRKAG3, RUNX1, SMAD4, STK3 and TBL1XR1) were significantly down-regulated in SQ and had a strong negative correlation with nniR-205, while one target gene (CDH3) was up-regulated in SQ and exhibited a strong positive correlation with miR-205.Conclusions We confirmed the high diagnostic accuracy of miR-205 in discriminating SQ from AC and SCLC in Chinese patients. Moreover, we identified 11 significant target genes of miR-205 which could be used for further functional studies as the basis for the development of SQ targeted therapies.