Department of Gerontology, Xiangya Hospital of Central South University, Changsha, Hunan 410078, P.R. China.
Mol Med Rep. 2017 Oct;16(4):5129-5136. doi: 10.3892/mmr.2017.7244. Epub 2017 Aug 14.
Non‑small‑cell lung cancer (NSCLC) is a leading cause of cancer mortality worldwide. The most common subtypes of NSCLC are adenocarcinoma (AC) and squamous cell carcinoma (SCC). However, the pathophysiological mechanisms contributing to AC and SCC are still largely unknown, especially the roles of long non‑coding RNAs (lncRNAs). The present study identified differentially expressed lncRNAs between lung AC and SCC by re‑annotation of NSCLC microarray data analysis profiling. The potential functions of lncRNAs were predicted by using coding‑non‑coding gene co‑expressing network. Reverse transcription-quantitative polymerase chain reaction (RT‑qPCR) was used to investigate lncRNA expression levels in AC cell lines (A549 and L78), SCC cell lines (H226 and H520) and normal cells (NL‑20). Western blotting analysis was used to investigate the protein expression levels in these cell lines. A total of 65 lncRNAs were differentially expressed between AC and SCC including 28 lncRNAs that were downregulated in SCC subtypes compared with those in AC ones, and 37 upregulated lncRNAs in SCC subtypes compared with AC subtypes. Three lncRNAs, sex determining region Y‑box 2 overlapping transcript (SOX2‑OT), NCBP2 antisense RNA 2 (NCBP2‑AS2) and ubiquitin like with PHD and ring finger domains 1 (UHRF1), were predicted to be associated with lung cancer; RT‑qPCR confirmed that SOX2‑OT and NCBP2‑AS2 were associated with lung cancer. Finally, western blot assays demonstrated that there was no difference in β‑catenin and glycogen synthase kinase 3β (GSK‑3β) expression in cancer cells compared with NL‑20, but increased phosphorylated (p‑)β‑catenin and p‑GSK‑3β was detected in lung cancer cell lines compared with NL‑20, particularly in A549 cells. Although these results require further experimental verification, the analysis of lncRNA signatures between AC and SCC has provided insights into the regulatory mechanism of NSCLC development.
非小细胞肺癌（NSCLC）是全球癌症死亡的主要原因。NSCLC 最常见的亚型是腺癌（AC）和鳞状细胞癌（SCC）。然而，导致 AC 和 SCC 的病理生理机制在很大程度上仍然未知，特别是长非编码 RNA（lncRNA）的作用。本研究通过 NSCLC 微阵列数据分析谱的重新注释，鉴定了肺 AC 和 SCC 之间差异表达的 lncRNA。通过编码-非编码基因共表达网络预测 lncRNA 的潜在功能。逆转录定量聚合酶链反应（RT-qPCR）用于研究 AC 细胞系（A549 和 L78）、SCC 细胞系（H226 和 H520）和正常细胞（NL-20）中 lncRNA 的表达水平。Western blot 分析用于研究这些细胞系中的蛋白表达水平。AC 和 SCC 之间共有 65 个 lncRNA 表达差异，其中 28 个 lncRNA 在 SCC 亚型中下调，37 个 lncRNA 在 SCC 亚型中上调。三个 lncRNA，性别决定区 Y 框 2 重叠转录物（SOX2-OT）、NCBP2 反义 RNA 2（NCBP2-AS2）和泛素样与 PH 和环指域 1（UHRF1），被预测与肺癌有关；RT-qPCR 证实 SOX2-OT 和 NCBP2-AS2 与肺癌有关。最后，Western blot 检测结果表明，与 NL-20 相比，癌细胞中β-连环蛋白和糖原合成酶激酶 3β（GSK-3β）的表达没有差异，但与 NL-20 相比，肺癌细胞系中检测到磷酸化（p）β-连环蛋白和 p-GSK-3β增加，特别是在 A549 细胞中。尽管这些结果需要进一步的实验验证，但对 AC 和 SCC 之间 lncRNA 特征的分析为 NSCLC 发展的调控机制提供了新的见解。
