Peng Kaipeng, Ren Xiangqing, Ren Qian
The First Clinical Medical College, Lanzhou University, Lanzhou, China.
Department of Gastroenterology, The First Hospital of Lanzhou University, Lanzhou, China.
Front Genet. 2022 Aug 25;13:888672. doi: 10.3389/fgene.2022.888672. eCollection 2022.
Gastric cancer (GC) is still notorious for its poor prognosis and aggressive characteristics. Though great developments have been made in diagnosis and therapy for GC, the prognosis of patient is still perishing. In this study, differentially expressed genes (DEGs) in GC were first screened using three Gene Expression Omnibus (GEO) datasets (GSE13911, GSE29998, and GSE26899). Second, The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) data were used to validate expression of these DEGs and perform survival analysis. We selected seven candidate genes (CAMK2N1, OLFML2B, AKR7A3, CYP4X1, FMO5, MT1H, and MT1X) to carry out the next analysis. To construct the ceRNA network, we screened the most potential upstream ncRNAs of the candidate genes. A series of bioinformatics analyses, including expression analysis, correlation analysis, and survival analysis, revealed that the SNHG10-hsa-miR-378a-3p might be the most potential regulatory axis in GC. Then, the expression of CAMK2N1, miR-378a-3p, and SNHG10 was verified in GC cell lines (GES-1, MGC-803, BGC-823, HGC-27, MKN-45, and AGS) by qRT-PCR and Western blotting. We found that SNHG10 and CAMK2N1 were highly expressed in gastric cancer lines, and the miR-378a-3p was lowly expressed in BGC-823, HGC-27, and MKN-45. Furthermore, CAMK2N1 levels were significantly negatively associated with tumor immune cell infiltration, biomarkers of immune cells, and immune checkpoint expression. In summary, our results suggest that the ncRNA-mediated high expression of CAMK2N1 is associated with poor prognosis and tumor immune infiltration of GC.
胃癌(GC)因其预后差和侵袭性特征仍然声名狼藉。尽管在胃癌的诊断和治疗方面取得了巨大进展,但患者的预后仍然不佳。在本研究中,首先使用三个基因表达综合数据库(GEO)数据集(GSE13911、GSE29998和GSE26899)筛选胃癌中的差异表达基因(DEG)。其次,利用癌症基因组图谱(TCGA)和基因型-组织表达(GTEx)数据验证这些DEG的表达并进行生存分析。我们选择了七个候选基因(CAMK2N1、OLFML2B、AKR7A3、CYP4X1、FMO5、MT1H和MT1X)进行下一步分析。为构建ceRNA网络,我们筛选了候选基因最具潜力的上游非编码RNA。一系列生物信息学分析,包括表达分析、相关性分析和生存分析,表明SNHG10-hsa-miR-378a-3p可能是胃癌中最具潜力的调控轴。然后,通过qRT-PCR和蛋白质印迹法在胃癌细胞系(GES-1、MGC-803、BGC-823、HGC-27、MKN-45和AGS)中验证CAMK2N1、miR-378a-3p和SNHG10的表达。我们发现SNHG10和CAMK2N1在胃癌细胞系中高表达,而miR-378a-3p在BGC-823、HGC-27和MKN-45中低表达。此外,CAMK2N1水平与肿瘤免疫细胞浸润、免疫细胞生物标志物和免疫检查点表达显著负相关。总之,我们的结果表明,ncRNA介导的CAMK2N1高表达与胃癌的不良预后和肿瘤免疫浸润有关。