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. 2002 Nov 4;87(10):1153-61.
doi: 10.1038/sj.bjc.6600580.

Differential gene expression profiles of gastric cancer cells established from primary tumour and malignant ascites

C Sakakura  1 , A Hagiwara, M Nakanishi, K Shimomura, T Takagi, R Yasuoka, Y Fujita, T Abe, Y Ichikawa, S Takahashi, T Ishikawa, I Nishizuka, T Morita, H Shimada, Y Okazaki, Y Hayashizaki, H Yamagishi
Affiliations

Differential gene expression profiles of gastric cancer cells established from primary tumour and malignant ascites

C Sakakura et al. Br J Cancer. .

Abstract

Advanced gastric cancer is often accompanied by metastasis to the peritoneum, resulting in a high mortality rate. Mechanisms involved in gastric cancer metastasis have not been fully clarified because metastasis involves multiple steps and requires a combination of altered expressions of many different genes. Thus, independent analysis of any single gene would be insufficient to understand all of the aspects of gastric cancer peritoneal dissemination. In this study, we performed a global analysis of the differential gene expression of a gastric cancer cell line established from a primary main tumour (SNU-1) and of other cell lines established from the metastasis to the peritoneal cavity (SNU-5, SNU-16, SNU-620, KATO-III and GT3TKB). The application of a high-density cDNA microarray method made it possible to analyse the expression of approximately 21 168 genes. Our examinations of SNU-5, SNU-16, SNU-620, KATO-III and GT3TKB showed that 24 genes were up-regulated and 17 genes down-regulated besides expression sequence tags. The analysis revealed the following altered expression such as: (a) up-regulation of CD44 (cell adhesion), keratins 7, 8, and 14 (epitherial marker), aldehyde dehydrogenase (drug metabolism), CD9 and IP3 receptor type3 (signal transduction); (b) down-regulation of IL2 receptor gamma, IL4-Stat (immune response), p27 (cell cycle) and integrin beta4 (adhesion) in gastric cancer cells from malignant ascites. We then analysed eight gastric cancer cell lines with Northern blot and observed preferential up-regulation and down-regulation of these selected genes in cells prone to peritoneal dissemination. Reverse transcriptase-polymerase chain reaction confirmed that several genes selected by DNA microarray were also overexpressed in clinical samples of malignant ascites. It is therefore considered that these genes may be related to the peritoneal dissemination of gastric cancers. The results of this global gene expression analysis of gastric cancer cells with peritoneal dissemination, promise to provide a new insight into the study of human gastric cancer peritoneal dissemination.

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Figures

Figure 1
Figure 1
High-density cDNA microarray analysis. Validation in expression of selected genes from 21 68 clones in six gastric cancer cell lines. Data are represented in a matrix format: each row represents a single gene, and each column an experimental sample. In each sample, the ratio of the number of transcripts of each gene to the medium abundance of the gene's transcript among all the cell lines is represented by the colour of the corresponding cell in the matrix. Green square,transcript level below the median; black squares, transcript levels equal to the medium; red squares, transcript levels greater than the median. Colour saturation reflects the magnitude of the ratio relative to the median for each set of samples. (A) Cluster analysis of selected genes of six gastric cancer cell lines using the RIKEN cDNA microarray. Bars to the right identify the location of the inserts displayed in panels B and C. (B) Up-regulated genes in gastric cancer cells from malignant ascites. (C) Down-regulated genes in gastric cancer cells from malignant ascites
Figure 2
Figure 2
Northern blot analysis of differently expressed genes in eight gastric cancer cells. Up-regulated genes in gastric cancer cells from malignant ascites in comparison to primary lesion
Figure 3
Figure 3
Northern blot analysis of differently expressed genes in eight gastric cancer cells. Down-regulated genes in gastric cancer cells from malignant ascites other than those from primary lesion
Figure 4
Figure 4
Expression of selected genes in clinical samples of peritoneal dissemination. Representative RT–PCR results of six peritoneal washes from patients with benign disease (cytology negative) and from four gastric cancer patients with malignant ascites (cytology positive). Arrows indicate the specific bands of each gene
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