Identification of differentially expressed genes in rat hepatoma cell lines using subtraction and microarray

Rat hepatoma Fao is a well-differentiated cell line that expresses numerous liver-specific enzymes and liver-enriched transcription factors, whereas the C2 cell line derived from Fao exhibits dedifferentiated phenotypes and fails to express these differentiated traits. Hence, to identify the gene related to the differentiation status in these cells, we analyzed differentially expressed genes between Fao and C2 cells using a microarray of 1,000 identified cDNA clones, and isolated a cDNA clone from subtractive library constructed by suppression subtractive hybridization. D6.1A, a member of the tetraspanin protein superfamily, was identified as a gene enhanced in C2 cells compared with Fao cells. Interestingly, expression of this gene was also induced following liver injury by CCl4 and in liver regeneration following partial hepatectomy. These results suggest that the D6.1A gene is related to stimulation of cell proliferation and differentiation status in C2 hepatoma cells.