Department of Toxicology, School of Public Health, China Medical University, Shenyang, Liaoning 110122, P.R. China.
Department of Thoracic Surgery Ward 2, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110122, P.R. China.
Int J Oncol. 2018 Jun;52(6):2155-2165. doi: 10.3892/ijo.2018.4347. Epub 2018 Mar 29.
Numerous genes are arranged in complex overlapping and interlaced patterns, and such arrangements potentially contribute to the regulation of gene expression. Previous studies have demonstrated that a region in chromosome 19q13.2-3 encompassing the overlapping genes excision repair cross-complementation group 1 (ERCC1), CD3e molecule associated protein (CD3EAP) and protein phosphatase 1 regulatory subunit 13 like (PPP1R13L) was found to be associated with the risk and prognosis of non-small cell lung cancer (NSCLC). The present study confirmed the hypothesis that there are co-expression patterns among these overlapping genes. The suggestive bioinformatic evidence of The Cancer Genome Atlas was verified by quantitative polymerase chain reaction (qPCR) analysis of NSCLC tissue samples. In addition, a cisplatin-induced DNA damage cell model was assessed by microarray analysis, qPCR and 3' rapid amplification of cDNA ends (3'RACE) to verify and quantify the expression levels of co-expressed alternative splicing isoforms in the NSCLC tissues, as well as in cancer A549 and normal 16HBE cells. The expression of CD3EAP exon 1 was demonstrated to be significantly associated with PPP1R13L exon 1, while CD3EAP exon 3 was significantly associated with ERCC1 exon 11 in normal and NSCLC tissues. It was observed that short transcripts of ERCC1, CD3EAP and PPP1R13L are co-expressed in A549 cells and full-length transcripts are co-expressed in 16HBE cells. Furthermore, a novel transcriptional regulation pattern was described based on the positional associations of overlapping genes. The region encompassing the overlapping genes ERCC1, CD3EAP and PPP1R13L may be involved in linking the upstream and downstream genes, while the different splicing isoforms of ERCC1 affect the expression of its overlapping genes, suggesting potential application in cisplatin resistance in NSCLC treatment.
大量基因以复杂的重叠和交错模式排列，这种排列方式可能有助于基因表达的调控。先前的研究表明，19q13.2-3 号染色体上包含重叠基因切除修复交叉互补组 1 (ERCC1)、CD3e 分子相关蛋白 (CD3EAP) 和蛋白磷酸酶 1 调节亚基 13 样 (PPP1R13L) 的区域与非小细胞肺癌 (NSCLC) 的风险和预后相关。本研究证实了这些重叠基因之间存在共表达模式的假设。通过对 NSCLC 组织样本进行定量聚合酶链反应 (qPCR) 分析，验证了癌症基因组图谱的提示性生物信息学证据。此外，通过微阵列分析、qPCR 和 3'快速扩增 cDNA 末端 (3'RACE) 评估顺铂诱导的 DNA 损伤细胞模型，以验证和量化 NSCLC 组织以及癌症 A549 和正常 16HBE 细胞中共同表达的选择性剪接异构体的表达水平。结果表明，CD3EAP 外显子 1 的表达与 PPP1R13L 外显子 1 显著相关，而正常和 NSCLC 组织中 CD3EAP 外显子 3 与 ERCC1 外显子 11 显著相关。观察到 A549 细胞中 ERCC1、CD3EAP 和 PPP1R13L 的短转录本共表达，而 16HBE 细胞中全长转录本共表达。此外，还根据重叠基因的位置关联描述了一种新的转录调控模式。包含重叠基因 ERCC1、CD3EAP 和 PPP1R13L 的区域可能参与连接上下游基因，而 ERCC1 的不同剪接异构体影响其重叠基因的表达，这表明其在 NSCLC 顺铂耐药治疗中有潜在应用。
