Department of Respiratory and Critical Care Medicine, The 900th Hospital of Joint Logistic Support Force, Fuzhou, Fujian, China.
Department of Thoracic Surgery, The 900th Hospital of Joint Logistic Support Force, Fuzhou, Fujian, China.
Mol Genet Genomic Med. 2020 Dec;8(12):e1521. doi: 10.1002/mgg3.1521. Epub 2020 Nov 6.
This study aimed to identify critical genes involved in the tumor biology of lung cancer via datamining of The Cancer Genome Atlas (TCGA) with special focus on gene copy number variation.
Genomic deletion and amplification were analyzed with cBioportal online tools. Relative expression of Cyclin Dependent Kinase Inhibitor 2A (CDKN2A) was analyzed by both real-time polymerase chain reaction (PCR) and Western blot. The abundance of methylthioadenosine phosphorylase (MTAP) and epithelial-mesenchymal transition markers were analyzed by real-time PCR. Cell proliferation was determined by cell counting kit-8 method and cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The cell migration and invasion were measured with transwell chamber assay, and migrative capacity was further evaluated by wound healing assay.
We found the frequent loss of CDKN2A was associated with its downregulation in lung cancer, and siRNA-mediated CDNKN2A knockdown significantly stimulated cell proliferation, invasion, and migration. Mechanistically, we unraveled that MTAP, which was positively correlated with CDKN2A, predominantly mediated the antitumoral function of CDKN2A in lung cancer.
Our study consolidated the involvement of CDKN2A-MTAP signaling in the context of lung cancer.
本研究通过对癌症基因组图谱（TCGA）的数据挖掘，特别关注基因拷贝数变异，旨在确定与肺癌肿瘤生物学相关的关键基因。
利用 cBioportal 在线工具分析基因组缺失和扩增。通过实时聚合酶链反应（PCR）和 Western blot 分析细胞周期蛋白依赖性激酶抑制剂 2A（CDKN2A）的相对表达。通过实时 PCR 分析甲基硫腺苷磷酸化酶（MTAP）和上皮-间充质转化标志物的丰度。通过细胞计数试剂盒-8 法测定细胞增殖，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测量细胞活力。通过 Transwell 室测定细胞迁移和侵袭，通过划痕愈合试验进一步评估迁移能力。
我们发现 CDKN2A 的频繁缺失与其在肺癌中的下调有关，siRNA 介导的 CDNKN2A 敲低显著刺激细胞增殖、侵袭和迁移。从机制上讲，我们揭示了与 CDKN2A 呈正相关的 MTAP 主要介导了 CDKN2A 在肺癌中的抗肿瘤功能。
我们的研究证实了 CDKN2A-MTAP 信号在肺癌中的参与。
