Wang Peng-Peng, Ding Si-Yu, Sun Yuan-Yuan, Li Yun-Hui, Fu Wei-Neng
Department of Medical Genetics, China Medical University, Shenyang, China.
Department of Laboratory Medicine, General Hospital of Northern Theater Command, Shenyang, China.
Front Oncol. 2021 Feb 18;10:564733. doi: 10.3389/fonc.2020.564733. eCollection 2020.
MYCT1, a target of c-Myc, inhibits laryngeal cancer cell migration, but the underlying mechanism remains unclear. In the study, we detected differentially expressed genes (DEGs) from laryngeal cancer cells transfected by MYCT1 using RNA-seq (GSE123275). DEGs from head and neck squamous cell carcinoma (HNSCC) were first screened by comparison of transcription data from the Gene Expression Omnibus (GSE6631) and the Cancer Genome Atlas (TCGA) datasets using weighted gene co-expression network analysis (WGCNA). GO and KEGG pathway analysis explained the functions of the DEGs. The DEGs overlapped between GSE6631and TCGA datasets were then compared with ours to find the key DEGs downstream of MYCT1 related to the adhesion and migration of laryngeal cancer cells. qRT-PCR and Western blot were applied to validate gene expression at mRNA and protein levels, respectively. Finally, the cell adhesion, migration, and wound healing assays were to check cell adhesion and migration abilities, respectively. As results, 39 overlapping genes were enriched in the GSE6631 and TCGA datasets, and most of them revealed adhesion function. Thirteen of 39 genes including COL6 members COL6A1, COL6A2, and COL6A3 were overlapped in GSE6631, TCGA, and GSE123275 datasets. Similar to our RNA-seq results, we confirmed that COL6 is a target of MYCT1 in laryngeal cancer cells. We also found that MYCT1 inhibited the adhesion and migration of laryngeal cancer cells via COL6. These indicate that COL6 is a potential target of MYCT1 and participates the adhesion and migration of laryngeal cancer cells, which provides an important clue for further study on how MYCT1 regulating COL6 in laryngeal cancer progression.
MYCT1是c-Myc的一个靶点,可抑制喉癌细胞迁移,但其潜在机制尚不清楚。在本研究中,我们使用RNA测序(GSE123275)检测了经MYCT1转染的喉癌细胞中差异表达基因(DEG)。通过使用加权基因共表达网络分析(WGCNA)比较来自基因表达综合数据库(GSE6631)和癌症基因组图谱(TCGA)数据集的转录数据,首先筛选了头颈部鳞状细胞癌(HNSCC)的DEG。基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析解释了DEG的功能。然后将GSE6631和TCGA数据集之间重叠的DEG与我们的数据进行比较,以找到与喉癌细胞黏附和迁移相关的MYCT1下游关键DEG。分别应用qRT-PCR和蛋白质印迹法在mRNA和蛋白质水平验证基因表达。最后,进行细胞黏附、迁移和伤口愈合试验,分别检查细胞黏附和迁移能力。结果显示,39个重叠基因在GSE6631和TCGA数据集中富集,其中大多数具有黏附功能。39个基因中的13个,包括胶原蛋白VI(COL6)成员COL6A1、COL6A2和COL6A3,在GSE6631、TCGA和GSE123275数据集中重叠。与我们的RNA测序结果相似,我们证实COL6是喉癌细胞中MYCT1的一个靶点。我们还发现MYCT1通过COL6抑制喉癌细胞的黏附和迁移。这些结果表明,COL6是MYCT1的一个潜在靶点,并参与喉癌细胞的黏附和迁移,这为进一步研究MYCT1如何在喉癌进展中调节COL6提供了重要线索。