Wei Jingsun, Xue Yiqi, Huo Xinying, Han Rongbo, Su Xinyu, Jin Yan, Zhao Wenjing, Chen Yuetong, Zhang Honghong, Dai Jiali, Chen Jinfei
Department of Oncology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China,
Cancer Manag Res. 2019 Feb 8;11:1275-1287. doi: 10.2147/CMAR.S186416. eCollection 2019.
Gastric hepatoid adenocarcinoma is a rare subtype of primary gastric cancer and is a high-grade form of malignancy. However, the pathogenesis and molecular biology of gastric hepatoid adenocarcinoma remain poorly understood. The aim of this study was to establish and characterize a new human gastric hepatoid adenocarcinoma cell line, GC-030-35.
The GC-030-35 cell line was established from tumor cells from a 58-year-old Chinese man with gastric hepatoid adenocarcinoma. The cultured cells underwent immunocytochemistry and flow cytometry to confirm the tumor cell phenotype. RNA sequencing was performed to analyze the differences in gene expression between GC-030-35 cells compared with normal gastric epithelial cells. A zebrafish assay was performed. Gene enrichment analysis and interrogation of the bioinformatics databases, the Gene Ontology (GO) database and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, were used for pathway analysis.
Flow cytometry analysis of the GC-030-35 cells showed a positive expression rate for CD44+ of 10.7%, high cell clonality, an average plating efficiency of 32%, cell-doubling time of 29.2 hours, and cell proliferation for >15 generations in serial culture. The zebrafish assay showed the ability of the GC-030-35 cells to proliferate, promote angiogenesis, and metastasize. RNA sequencing identified the functional clustering of 6,601 differentially expressed genes of GC-030-35, which were significantly different when compared with nonneoplastic gastric epithelial cells. Pathway enrichment analysis and interrogation of the GO and KEGG bioinformatics databases identified genes for microbial metabolism in diverse environments (63 genes), metabolism of xenobiotics by cytochrome P450 (CYP450; 25 genes), and the drug metabolism cytochrome P450 (28 genes).
A human gastric hepatoid adenocarcinoma cell line, GC-030-35, was developed and characterized by comparison with normal gastric epithelial cells. Bioinformatics and gene analysis data showed that the CYP450 gene was significantly differentially expressed by GC-030-35 cells.
胃肝样腺癌是原发性胃癌的一种罕见亚型，是一种高级别恶性肿瘤。然而，胃肝样腺癌的发病机制和分子生物学仍知之甚少。本研究的目的是建立并鉴定一种新的人胃肝样腺癌细胞系GC-030-35。
GC-030-35细胞系源自一名58岁患胃肝样腺癌的中国男性的肿瘤细胞。对培养的细胞进行免疫细胞化学和流式细胞术检测，以确认肿瘤细胞表型。进行RNA测序，分析GC-030-35细胞与正常胃上皮细胞之间基因表达的差异。开展斑马鱼试验。利用基因富集分析并查询生物信息学数据库，即基因本体论（GO）数据库和京都基因与基因组百科全书（KEGG）数据库，进行通路分析。
对GC-030-35细胞进行流式细胞术分析显示，CD44+阳性表达率为10.7%，细胞克隆性高，平均接种效率为32%，细胞倍增时间为29.2小时，且在连续培养中细胞增殖超过15代。斑马鱼试验显示GC-030-35细胞具有增殖、促进血管生成和转移的能力。RNA测序确定了GC-030-35的6601个差异表达基因的功能聚类，与非肿瘤性胃上皮细胞相比有显著差异。通路富集分析以及对GO和KEGG生物信息学数据库的查询确定了参与不同环境中微生物代谢的基因（63个基因）、细胞色素P450（CYP450）对外源生物的代谢（25个基因）以及药物代谢细胞色素P450（28个基因）。
建立了人胃肝样腺癌细胞系GC-030-35，并与正常胃上皮细胞进行比较以对其进行鉴定。生物信息学和基因分析数据表明，GC-030-35细胞中CYP450基因有显著差异表达。
