Establishment and characterization of the GC-030-35 cell line derived from gastric hepatoid adenocarcinoma

Abstract

Jingsun Wei,* Yiqi Xue,* Xinying Huo, Rongbo Han, Xinyu Su, Yan Jin, Wenjing Zhao, Yuetong Chen, Honghong Zhang, Jiali Dai, Jinfei Chen Department of Oncology, Nanjing First Hospital, Nanjing Medical University, Nanjing, China *These authors contributed equally to this work Purpose: 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.Materials and methods: 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.Results: 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).Conclusion: 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. Keywords: gastric hepatoid adenocarcinoma, cell line, differentially expressed genes, bioinformatics, carcinogenesis, cytochrome P450

Keywords

• gastric hepatoid adenocarcinoma
• cell line
• differentially expressed genes
• bioinformatics
• carcinogenesis
• cytochrome P450.