Zhai Jingming, Lyu Ting, Guo Yimin, An Yanhui, Xiang Yali, Xie Linying, Zeng Chao
Department of General Surgery, The First Affiliated Hospital and College of Clinical Medicine, Henan University of Science and Technology, Luoyang, China.
Department of Pathology, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shenzhen, China.
Sci Rep. 2025 May 9;15(1):16146. doi: 10.1038/s41598-025-99632-2.
Gastric cancer of young adults is poorly differentiated and has a poor prognosis. However, there are few reports regarding the genetic alteration in gastric cancer of young adults. Bioinformatics methods were used to screen the key genes and signaling pathways of gastric cancer in young adults, and molecular biology techniques were used to verify the key proteins involved in the occurrence and development of gastric cancer in young adults. RNA expression profile microarray data of gastric cancer patients ≤ 45 years old and > 45 years old were downloaded from the TCGA database, and differentially expressed genes were screened by Limma package. GO analysis and KEGG enrichment analysis of DEG were performed in the Gene Function annotation database (DAVID). CytoHubba is used to construct protein interaction networks (PPI) and perform visual analysis to screen out core genes. We obtained 10 hub molecules, including FBXO44, FBXO6, HERC5, FBXL13, FBXO41, NT5E, BMP4, TRIM36, ACAN, ALPL by PPI network with MCODE. Ingenuity Pathway Analysis predicts TBX1, DFNB31, TGFBR3, FBXO44, SLC7A8, DNM1, KITLG, MSH5, MLLT3, DUSP5, ADAMTSL4, ACPP and TBX1 as the transcription factors directly regulated by OTX2. OTX2 had the highest positive expression rate in gastric cancer of young adults by immunohistochemistry. Interference with OTX2 expression inhibits proliferation, migration, invasion and promotes differentiation, apoptosis of NUGC-4 cells (from 35 year old female). Moreover, after interfering OTX2 expression, the downstream proteins and signaling Pathways of OTX2 in NUGC-4 were further analyzed by Transcriptome sequencing and Ingenuity Pathways Analysis. We found interference with OTX2 expression inhibits CEBPB expression and activates calcitriol by IPA analysis, thereby promoting differentiation of NUGC-4. Therefore, OTX2 plays important roles in restraining the differentiation and promoting progression of gastric cancer cells in young adults. Moreover, OTX2/CEBPB signal axis is likely to be a key molecular event in regulating the differentiation of gastric cancer cells in young adults.
青年胃癌分化程度低，预后较差。然而，关于青年胃癌基因改变的报道较少。本研究运用生物信息学方法筛选青年胃癌关键基因及信号通路，并采用分子生物学技术验证参与青年胃癌发生发展的关键蛋白。从TCGA数据库下载年龄≤45岁和＞45岁胃癌患者的RNA表达谱芯片数据，用Limma软件包筛选差异表达基因。在基因功能注释数据库（DAVID）中对差异表达基因进行GO分析和KEGG富集分析。利用CytoHubba构建蛋白质相互作用网络（PPI）并进行可视化分析以筛选核心基因。通过PPI网络结合MCODE，我们获得了10个枢纽分子，包括FBXO44、FBXO6、HERC5、FBXL13、FBXO41、NT5E、BMP4、TRIM36、ACAN、ALPL。 Ingenuity通路分析预测TBX1、DFNB31、TGFBR3、FBXO44、SLC7A8、DNM1、KITLG、MSH5、MLLT3、DUSP5、ADAMTSL4、ACPP和TBX1为受OTX2直接调控的转录因子。免疫组化结果显示OTX2在青年胃癌中阳性表达率最高。干扰OTX2表达可抑制NUGC-4细胞（来自35岁女性）的增殖、迁移、侵袭，并促进其分化、凋亡。此外，干扰OTX2表达后，通过转录组测序和 Ingenuity通路分析进一步分析了OTX2在NUGC-4中的下游蛋白和信号通路。我们发现干扰OTX2表达可抑制CEBPB表达，并通过IPA分析激活骨化三醇，从而促进NUGC-4的分化。因此，OTX2在抑制青年胃癌细胞分化和促进其进展中起重要作用。此外，OTX2/CEBPB信号轴可能是调节青年胃癌细胞分化的关键分子事件。
