Differential gene expression between asymptomatic HBV carriers and normal adults.
BACKGROUND
There are more than 100 million asymptomatic HBV carriers (ASCs) in China and they are at a high risk of developing liver disease, which creates a serious health problem. But more than 90% of normal adults clear virus from primary HBV infection, so the difference of gene expression between ASCs and normal adults determines the differential outcomes. To identify differentially expressed genes in ASCs compared to normal adults, we used suppression subtractive hybridization to compare gene expression.


METHODS
cDNA subtracted libraries were constructed by suppression subtracted hybridization from peripheral blood monocytes of ASCs and normal adults, the subtracted clones were prescreened by dot blot hybridization, and the levels of genes of interest were confirmed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR).


RESULTS
One hundred and two and 96 positive clones were acquired from the A-N and N-A libraries, respectively, and 14 and 11 clones were identified by dot blot hybridization in the A-N and N-A libraries. Two genes were confirmed as differentially expressed between a set of ASC and normal adult samples by real-time RT-PCR.


CONCLUSIONS
Differentially expressed genes in peripheral blood mononuclear cells between ASCs and normal adults were isolated by suppression subtractive hybridization, and included some new genes. Of the upregulated genes in ASCs, checkpoint suppressor 1 is associated with DNA damage-induced cell cycle arrest. Perforin 1 is associated with inflammation. The information about such alterations in gene expression could be useful for elucidating the mechanisms of ASC pathogenesis.
Tang
Cui-Lan
CL
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Chen
Zhi
Z
D000328:Adult; D002353:Carrier State; D016022:Case-Control Studies; D018797:Cell Cycle Proteins / Q000235:genetics; D018450:Disease Progression; D005260:Female; D051858:Forkhead Transcription Factors; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D015723:Gene Library; D020022:Genetic Predisposition to Disease; D006509:Hepatitis B / Q000235:genetics*; D006801:Humans; D008297:Male; D009693:Nucleic Acid Hybridization; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018797:Cell Cycle Proteins; C106158:FOXN3 protein, human; D051858:Forkhead Transcription Factors; C510468:PRF1 protein, human; D052899:Pore Forming Cytotoxic Proteins; D012097:Repressor Proteins; D054353:Perforin