Screening and
Identification of Gastric Adenocarcinoma Metastasis-related Genes by Using cDNA
Microarray Coupled to FDD-PCR
WANG Jian-Hua, CHEN Shi-Shu*
( Laboratory of Molecular
Biology, Research Center for Human Gene Therapy, Shanghai Second Medical
University, Shanghai 200025, China )
Abstract To clone
gastric adenocarcinoma metastasis related genes, RF-1 cell line (primary tumor
of a gastric adenocarcinoma patient ) and RF-48 cell line (its metastatic
counterpart) were used as a model for studying the molecular mechanism of tumor
metastasis. Two fluorescent cDNA probes, labeled with Cy3 and Cy5 dyes, were
prepared from RF-1 and RF-48 mRNA samples by reverse transcription method. The
two color probes were then mixed and hybridized to the cDNA chip constructed by
double-dots of 4 096 human genes, and scanned at two wavelengths. The
experiment was repeated for 2 times. Differential expression genes from the
above two cells were analyzed using the computer. 138 in all genes (3.4%)
revealed differential expression in RF-48 cells compared with RF-1 cells:
81(2.1%) genes revealed apparent up-regulation, and 56(1.3%) genes revealed
down-regulation. 45 genes involved in gastric adenocarcinoma metastasis were
cloned using fluorescent differential display-PCR (FDD-PCR), including 3 novel
genes. There were 7 differential expression genes that agreed with each other
in two detection methods. The possible roles of some differential expressed
genes, which maybe involved in the mechanism of tumor metastasis, were
discussed. cDNA chip was used to analyze gene expression in a high-throughput
and large scale manner, in combination with FDD-PCR for cloning unknown novel
genes. In conclusion, some genes related to metastasis were preliminarily
scanned, which would contribute to disclose the molecular mechanism of gastric
adenocracinoma metastasis.
Key words cDNA chip; fluorescent differential
display-PCR; gastric adenocracinoma;
metastasis-related genes
*Corresponding author: Tel,
86-21-63846592-776452; Fax, 86-21-34060742; e-mail, [email protected]