基于转录组测序的胃癌关键基因识别与影响机制分析

Indentification of key genes in gastric cancer and analysis of their influencing mechanisms based on transcriptome sequencing

摘要

目的：基于转录组测序（RNA-Seq）技术筛选胃癌（GC）进展关键基因，并推测其影响GC进展的潜在机制。方法：收集晚期GC腹膜转移患者行腹腔热灌注化疗（HIPEC）治疗前后大网膜转移结节进行高通量RNA测序，对差异表达基因进行GO和KEGG富集分析；qRT-PCR检测上调倍数最大的前10个基因在治疗前后的mRNA变化；通过siRNA技术抑制HGC27细胞系中关键基因A1CF的表达，采用MTT实验、细胞克隆形成实验及Transwell实验检测A1CF对GC细胞增殖和侵袭能力的影响。结果：HIPEC后，共有2087个基因上调，875个基因下调。差异基因主要功能富集在MAPK通路、NOD样受体通路和TNF通路等。在上调倍数最大的10个基因中，治疗后FOSB、DEFA5、EGR3、A1CF的mRNA水平显著上升。细胞实验显示，抑制关键基因A1CF后GC细胞的增殖和侵袭能力明显减弱（P＜0.05）。结论：FOSB、DEFA5、EGR3、A1CF是GC进展的关键基因，A1CF能够驱动GC细胞增殖与侵袭过程，促进GC进展。

Abstract

Objective: To delineate the differential gene expression profiles in patients with advanced gastric cancer(GC) and peritoneal metastasis before and after hyperthermic intraperitoneal chemotherapy(HIPEC) using RNA sequencing(RNA-Seq), thereby identifying pivotal genes implicated in GC progression and elucidating the mechanistic role of A1CF in promoting GC cell proliferation and invasion, so as to provide new potential targeted therapy strategies for advanced GC patients. Methods: Omentum metastatic nodules were collected from advanced GC patients with peritoneal metastasis pre- and post-HIPEC for RNA-Seq analysis. Differentially expressed genes(DEGs) were subjected to Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses. The top 10 most upregulated genes were validated via qRT-PCR to assess mRNA expression changes. A1CF expression was silenced in HGC27 cells using siRNA, followed by functional assays including MTT, colony formation, and transwell experiments to evaluate its impact on GC cell proliferation and invasion. Results: HIPEC treatment induced significant alterations in gene expression, with 2,087 genes upregulated and 875 genes downregulated. DEGs were predominantly enriched in pathways including MAPK signaling, NOD-like receptor signaling, and TNF signaling. Among the top 10 upregulated genes, FOSB, DEFA5, EGR3, and A1CF exhibited markedly elevated mRNA levels post-treatment. Functional inhibition of A1CF substantially attenuated the proliferative and invasive capacities of GC cells(P＜0.05). Conclusion: FOSB, DEFA5, EGR3, and A1CF are key genes in the progression of gastric cancer(GC), and A1CF can drive the proliferation and invasion of GC cells to promote the progression of GC.

关键词

胃肿瘤 / 腹腔热灌注化疗 / RNA测序 / A1CF

Key words

Gastric neoplasms / Intraperitoneal hyperthermic chemotherapy / RNA sequencing / A1CF

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基金资助

国家自然科学基金资助项目(82073271)