Research Center of Medical Sciences, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China.
Department of Otolaryngology & Head and Neck, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China.
Biomed Res Int. 2019 Jun 11;2019:7246491. doi: 10.1155/2019/7246491. eCollection 2019.
Our previous studies showed that ZBTB7A played an important role in promoting nasopharyngeal carcinoma (NPC) progression. However, molecular mechanisms of different levels of ZBTB7A are still unclear. It is necessary to search molecular markers which are closely connected with ZBTB7A. We selected NPC sublines CNE2 with stably transfecting empty plasmid (negative control, NC) and short hair RNA (shRNA) plasmid targeting ZBTB7A as research objectives. Microarray was used to screen differentially expressed long noncoding RNAs (lncRNAs) and messenger RNAs (mRNAs) via shRNA-CNE2 versus NC-CNE2. Quantitative PCR (qPCR) was used to validate lncRNAs and mRNAs from the sublines, chronic rhinitis, and NPC tissues. Bioinformatics was used to analyze regulatory pathways which were connected with ZBTB7A. The 1501 lncRNAs (long noncoding RNAs) and 1275 differentially expressed mRNAs were upregulated or downregulated over 2-fold. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the upregulated or downregulated carbohydrate and lipid metabolisms probably involved in carcinogenicity of shRNA-CNE2 (P-value cut-off was 0.05). In order to find the molecular mechanisms of ZBTB7A, we validated 12 differentially expressed lncRNAs and their nearby mRNAs by qPCR. Most of the differentially expressed mRNAs are closely connected with carbohydrate and lipid metabolisms in multiply cancers. Furthermore, part of them were validated in NPC and rhinitis tissues by qPCR. As a result, NR_047538, ENST00000442852, and fatty acid synthase (FASN) were closely associated with NPC. ZBTB7A had a positive association with NR_047538 and negative associations with ENST00000442852 and FASN. The results probably provide novel candidate biomarkers for NPC progression with different levels of ZBTB7A.
我们之前的研究表明,ZBTB7A 在促进鼻咽癌(NPC)进展中发挥着重要作用。然而,不同水平的 ZBTB7A 的分子机制仍不清楚。有必要寻找与 ZBTB7A 密切相关的分子标志物。我们选择了稳定转染空质粒(阴性对照,NC)和靶向 ZBTB7A 的短发夹 RNA(shRNA)质粒的 NPC 亚系 CNE2 作为研究对象。通过 shRNA-CNE2 与 NC-CNE2 的比较,使用微阵列筛选差异表达的长链非编码 RNA(lncRNA)和信使 RNA(mRNA)。使用定量 PCR(qPCR)验证亚系、慢性鼻炎和 NPC 组织中的 lncRNA 和 mRNA。使用生物信息学分析与 ZBTB7A 相关的调节途径。上调或下调超过 2 倍的 1501 个 lncRNA(长链非编码 RNA)和 1275 个差异表达的 mRNA。基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路分析表明,上调或下调的碳水化合物和脂质代谢可能与 shRNA-CNE2 的致癌性有关(P 值截止值为 0.05)。为了寻找 ZBTB7A 的分子机制,我们通过 qPCR 验证了 12 个差异表达的 lncRNA 及其附近的 mRNA。大多数差异表达的 mRNA 与多种癌症中的碳水化合物和脂质代谢密切相关。此外,部分在 NPC 和鼻炎组织中通过 qPCR 进行了验证。结果表明,NR_047538、ENST00000442852 和脂肪酸合酶(FASN)与 NPC 密切相关。ZBTB7A 与 NR_047538 呈正相关,与 ENST00000442852 和 FASN 呈负相关。这些结果可能为不同水平的 ZBTB7A 提供 NPC 进展的新候选生物标志物。