[Gene expression profiles and proteomics analysis of the cell transfected with the mutant type of COOH-terminal deleted of hepatitis B virus X].
OBJECTIVE
To investigate the mechanism of the different biological impacts of HBx3'-40, an engineered deletion mutant lacking the last 40 C-terminal amino acids.


METHODS
Human hepatocellular cells of the line Huh7 were transfected with HBx3'-40 or wtHBx (wild type HBx) constructs. An oligo cDNA microarray containing 21074 human genes and Ests was utilized to examine the different gene expression between HBx3'-40 and paired control wtHBx cells. A series of methods, including immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2-DE), silver staining, and PDQuest 2-D analysis software were used to analyze the differential protein-spots between HBx3'-40 and wtHBx cells. Selected differential protein-spots were identified with peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-fight mass spectrometry (MALDI-TOF-MS) and database searching.


RESULTS
165 (0.82%) candidate genes showing aberrant expression under HBx3'-40 induction were identified, of which 144 were up-regulated while 21 were down-regulated. Compared with wtHBx group, there were 135 +/- 13 differently protein spots in HBx3'-40 group by 2-DE. Of them, 7 significantly different protein spots were identified using mass spectrometry and computer matching with protein database. Most of the differently expressed genes and proteins were involved in transcription, oncogenes and tumor suppressor genes or protein, cell adhesion, signal transduction pathways, metabolisms, etc.


CONCLUSIONS
The mutant HBx affects cell genes and proteins involved in various processes, especially in regulation of liver metabolism.
Liu
Xiao-hong
XH
Department of Pathology, Changhai Hospital, Shanghai 200433, China.
Chen
Ying
Y
Wang
Li
L
Zhao
Hai-hua
HH
Zhu
Ming-hua
MH
D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D017353:Gene Deletion; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D050505:Mutant Proteins / Q000235:genetics / Q000502:physiology; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology; D054334:Viral Regulatory and Accessory Proteins
D050505:Mutant Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein