1998 Fiscal Year Final Research Report Summary
Function of Hepatitis B virus X gene in carcinogenesis of liven cells
| Project/Area Number |
09670539
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Mie University |
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Principal Investigator |
TAKASE koujiro Mie University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (00163204)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Masaaki Mie University, Hospital, Lecturer, 医学部・附属病院, 講師 (00223181)
SHIMIZU Atsuya Mie University, Faculty of Medicine, Assistant, 医学部, 助手 (10283524)
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| Project Period (FY) |
1997 – 1998
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| Keywords | HBx / LBP / Two Hybrid System / Apoptosis |
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| Research Abstract |
The X gene lesion of HBV from nine patients with hepatocellular carcinoma were amplified by PCR and sequenced. The X gene sequences were well conserved and no point mutations were detected compared to these form the healthy HBV carriers.
The laminin binding protein (LBP) was indentified as a protein interacting with X protein in the yeast two hybrid system. The interaction was confirmed by far western blot using recombinant GST- fusion proteins. As LBP was reported to have function in metastasis and proliferation of cancer cells via laminin receptor, we will continue to investigate the functions of these interaction in carcinogenesis of liver cells.
A specific antibody which could recognize X protein was developed and purified. The specificity of this antibody was confirmed by the evidence that this antibody could detect a single band of X protein expressed in both E. Coil (DH5α,45 kDa GST-fusion protein) and cultured cells (using pcDNA vector).
The X protein expressed in cultured cells transfected by pcDNA3.1(-)/Myc-His vector was stained mainly in the cytoplasm with granular pattern using both anti-X and anti-Myc antibodies. The expressed X protein was also observed in nucleus.
The X protein positive cells were diffusely observed in the liver tissues (biopsy form the patients with chronic active hepatitis B) and clustering positive cells were seen more frequently around the portal area.
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