Investigating the Role of Hbx In the Development Of Hepatocellular Carcinoma

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.