Construction of a subtracted cDNA library of differentially expressed genes in human normal liver tissue and liver cirrhosis with primary hepatocellular carcinoma background

Objective To study the differentially expressed genes in cirrhosis compared with normal liver and make the differentially expressed cDNAs subtracted library. Methods The differentially expressed fragments were obtained by using of suppression subtractive hybridization(SSH), then T/A cloning was accomplished to construct the cDNA library of differentially expressed genes. Results Seventy positive bacteria clones were randomly picked and identified using PCR and enzyme restriction method. Sixty-five contain positive insert fragments which range from 200 to 2 000 bp. Conclusion A subtracted cDNA library of differentially expressed genes in human normal liver tissue and HCC tissue is constructed successfully with SSH and TLA cloning techniques. The experimental findings demonstrated that there have been some gene changes in liver cirrhosis stage which may affect or promote the liver carcinogenesis.