胃癌(GC)仍然因其不良预后和侵袭性特征而臭名昭著。尽管GC的诊断和治疗有了很大的发展,患者的预后仍在恶化。在这项研究中,首先使用三个基因表达综合(GEO)数据集(GSE13911、GSE29998和GSE26899)筛选GC中的差异表达基因(DEGs)。第二,癌症基因组图谱(TCGA)和基因型-组织表达(GTEx)数据用于验证这些DEGs的表达并进行存活分析。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。为了构建ceRNA网络,我们筛选了候选基因中最有潜力的上游ncRNAs。一系列生物信息学分析,包括表达分析,相关分析,和生存分析,揭示SNHG10-hsa-miR-378a-3p可能是GC中最潜在的调节轴。然后,CAMK2N1,miR-378a-3p,通过qRT-PCR和Western印迹在GC细胞系(GES-1,MGC-803,BGC-823,HGC-27,MKN-45和AGS)中验证了SNHG10。我们发现SNHG10和CAMK2N1在胃癌细胞系中高表达,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呈显著负相关,免疫细胞的生物标志物,和免疫检查点表达。总之,我们的结果表明,ncRNA介导的CAMK2N1高表达与GC的不良预后和肿瘤免疫浸润有关。
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.