Abstract:
Hepatocellular carcinoma (HCC) is a primary malignancy of the liver and occurs
predominantly in patients with underlying chronic liver disease and cirrhosis. The
carcinogenesis related to HCC is multifactorial and multi-step process and chronic
infection with HBV is considered as one of the etiological agent for the hepatocellular
carcinoma (besides many others). Currently many researches have supported the role of
Hepatitis B virus X gene and protein (HBx) in the development of HCC. HBx induced
carcinogenesis is a result of modulation of various signalling pathways and factors, such
as activation of various transcription factors e.g. NF-κB or interaction with cellular
oncogenes. This study aims at amplification of HBx sequences from patient samples,
cloning poten tionally important X sequence for downstream processes and mammalian
expression and transfection of cloned X sequence into HCC cell line. So, the study was
designed to screen the HBx protein from blood sera samples of the patients, HBx being
cloned in the Ptz57R vector, isolation of plasmid, and the transfection studies were carried
out for mammalian expression and also to investigate the role of HBx in the development
of HCC through transfection studies. The HCC cells were transfected by the HBx gene
cloned in the plasmid vector (pTZ57R/T) and the results indicated the presence of mRNA
of HBx in the transfected Huh7 cells as compared to controlled cells. The experiment was
successful and HBx gene (cloned in vector) was successfully transfected in HCC cell line.
Conclusively, the experiment was successful in amplifying HBx gene from selected
patients and the HBx gene (cloned in plasmid) was successfully transfected in HCC cell
line. The study will help in studying the role of HBx protein in HCC cell line and
development of HCC by transfecting normal liver cells with HBx, better understanding
of HBx (HBV etiology) in hepatocellular carcinoma will help in identification and
development of potential biomarkers for early diagnosis and treatment of HCC, this will
not only help in designing new strategies for the treatment of HCC but also in
identification of novel proteins for better understanding of HBx induced HCC.
