[Mechanism of hepatocyte transformation by HCV NS3 using two-dimensional electrophoresis and mass spectrometry].
OBJECTIVE
To investigate the proteome of hepatocyte transformation by hepatitis C virus (HCV) nonstructural protein 3 (NS3).


METHODS
Human hepatocyte line QSG7701 stably expressing HCV NS3 C-terminal deleted protein was constructed, which was named pRcHCNS3/QSG. Two-dimensional electrophoresis (2-DE) was used to separate the total protein of pRcHCNS3/QSG and pRcCMV transfected cells (pRcCMV/QSG) respectively. Differentially expressed protein spots were identified by mass spectrometry. Western blot confirmed the differentially expressed proteins.


RESULTS
2-DE profiles with high resolution and reproducibility were obtained. The average spots of pRcHCNS3/QSG and pRcCMV/QSG were (1183+/-77) and (1095+/-82) respectively, and (920+/-60) spots were matched. Twenty-one differentially expressed protein spots were chosen randomly and 15 were identified by mass spectrometry. Some proteins such as Ras, P38 and HD53 which were involved in signal transduction were increased in pRcHCNS3/QSG cells. Western blot also showed strong expression of phosphorylated P44/42 and P38 in pRcHCNS3/QSG cells. Other differentially expressed proteins were related to cell cycle regulation, immunoreaction, tumor invasion and metastasis, and liver metabolizability.


CONCLUSIONS
HCV NS3 might be involved in cell malignant transformation through affecting protein expression and signal transduction such as MAPK cascade. Further study on the signal transductions and their relationship would not only be helpful to explore the mechanism of HCV related HCC, but also provide a new idea for the molecular treatment of HCC.
He
Qiong-qiong
QQ
Department of Pathology, School of Basic Medical, Central South University, Changsha 410078, China.
Chen
Zhu-chu
ZC
Xiao
Xu-xian
XX
Feng
De-yun
DY
Xiao
Zhi-qiang
ZQ
Li
Cui
C
Li
Bo
B
Zhang
Peng-fei
PF
Cheng
Rui-xue
RX
D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D020543:Proteome; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases