Research Article

Screening and identification of peritoneal metastasis-related genes of gastric adenocarcinoma using a cDNA microarray

Published: June 25, 2012
Genet. Mol. Res. 11 (2) : 1682-1689 DOI: 10.4238/2012.June.25.1

Abstract

With the aim of identifying peritoneal metastasis-related genes in gastric cancer, we performed a broad analysis of differential gene expression between the parental cell line GC9811 and its highly metastatic peritoneal counterpart, cell line GC9811-P. Two fluorescent cDNA probes, labeled with Cy3 and Cy5 dyes, were prepared from GC9811 and GC9811-P mRNA samples by the reverse transcription method. The two color probes were then mixed and hybridized to a cDNA chip constructed with double-dots from 11,901 human genes; this was scanned at two wavelengths. The experiment was repeated twice. In GC9811-P cells, 218 genes were upregulated and 30 genes were downregulated compared with the parental cell lines. Some selected genes were confirmed by RT-PCR and Western blot; we found that S100A4 and CTNNB1 were upregulated and PTEN was downregulated in GC9811-P cells. Identification of these differentially expressed genes could contribute to disclose the molecular mechanisms involved and provide new targets for therapeutic intervention to avoid peritoneal dissemination of gastric adenocarcinoma.

With the aim of identifying peritoneal metastasis-related genes in gastric cancer, we performed a broad analysis of differential gene expression between the parental cell line GC9811 and its highly metastatic peritoneal counterpart, cell line GC9811-P. Two fluorescent cDNA probes, labeled with Cy3 and Cy5 dyes, were prepared from GC9811 and GC9811-P mRNA samples by the reverse transcription method. The two color probes were then mixed and hybridized to a cDNA chip constructed with double-dots from 11,901 human genes; this was scanned at two wavelengths. The experiment was repeated twice. In GC9811-P cells, 218 genes were upregulated and 30 genes were downregulated compared with the parental cell lines. Some selected genes were confirmed by RT-PCR and Western blot; we found that S100A4 and CTNNB1 were upregulated and PTEN was downregulated in GC9811-P cells. Identification of these differentially expressed genes could contribute to disclose the molecular mechanisms involved and provide new targets for therapeutic intervention to avoid peritoneal dissemination of gastric adenocarcinoma.