CAL-27 细胞对过表达 ZNF750 基因反应的 RNA 测序分析显示，细胞周期受到广泛调控。
RNA sequencing analysis of the CAL-27 cell response to over-expressed ZNF750 gene revealed an extensive regulation on cell cycle.
机构信息
Department of Oral and Maxillofacial Surgery, Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China.
Central laboratory of Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China.
出版信息
Biomed Pharmacother. 2019 Oct;118:109377. doi: 10.1016/j.biopha.2019.109377. Epub 2019 Aug 28.
BACKGROUND
Zinc-finger protein 750 (ZNF750) is a potential tumor suppressor in oral squamous cell carcinoma (OSCC). However, the molecular mechanisms underlying its anti-tumor effect remain elusive in OSCC. This study aimed to elucidate the genes and pathways involved in tumor suppression following the ZNF750 over-expression in OSCC cell line CAL-27 cell by using the genomics approach.
METHODS
The RNA sequence libraries were constructed, and the data were analyzed to identify differentially expressed genes (DEGs) between vector groups and ZNF750 groups (over-expressed ZNF750 in CAL-27 cell). QPCR and western-blot was used to validate differential expression of candidate genes with cell cycle regulation. The cell cycle distribution was analyzed by BrdU staining.
RESULTS
By RNA sequencing profiling, 7,131 genes were differentially expressed in ZNF750 groups. Among the DEGs, 3,285 genes were upregulated, 3,846 genes were downregulated and 4,507 genes were identified in three main categories (cellular_component, biological process and molecular function) based on the gene ontology (GO) classification. The Kyoto Encyclopedia of Genes and Genome (KEGG) pathway analysis defined the DEGs could be categorized into 280 pathways and identified the top two most significant pathways involved in spliceosome and cell cycle. Functional categorization and enrichment analysis revealed that most of DEGs involved in binding and catalytic activity, and the cell cycle associated genes were significantly enriched in response to ZNF750 over-expression. ZNF750 induced cell cycle arrest in G0/G1 phase of the cell cycle.
CONCLUSION
Data from this study revealed that the cell cycle pathway was a key factor involved in the anti-tumor effect of ZNF750 in CAL-27 cells.
背景
锌指蛋白 750（ZNF750）是口腔鳞状细胞癌（OSCC）中的一种潜在的肿瘤抑制因子。然而，其在 OSCC 中抗肿瘤作用的分子机制仍不清楚。本研究旨在通过基因组学方法阐明 ZNF750 在 CAL-27 细胞系中过表达后肿瘤抑制涉及的基因和途径。
方法
构建 RNA 序列文库，并对数据进行分析，以鉴定载体组和 ZNF750 组（CAL-27 细胞中过表达 ZNF750）之间差异表达的基因（DEGs）。使用 QPCR 和 Western blot 验证候选基因与细胞周期调控的差异表达。通过 BrdU 染色分析细胞周期分布。
结果
通过 RNA 测序分析，ZNF750 组中有 7131 个基因差异表达。在 DEGs 中，上调的基因有 3285 个，下调的基因有 3846 个，根据基因本体（GO）分类，有 4507 个基因被确定为三个主要类别（细胞成分、生物过程和分子功能）。京都基因与基因组百科全书（KEGG）途径分析定义 DEGs 可分为 280 条途径，并确定了参与剪接体和细胞周期的两个最重要的途径。功能分类和富集分析表明，大多数 DEGs 参与结合和催化活性，与细胞周期相关的基因在响应 ZNF750 过表达时显著富集。ZNF750 诱导细胞周期停滞在 G0/G1 期。
结论
本研究的数据表明，细胞周期途径是 ZNF750 在 CAL-27 细胞中抗肿瘤作用的关键因素。
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