PENGARUH PROTEIN X MUTAN T118N VIRUS HEPATITIS B TERHADAP EKSPRESI BCL-2 PADA SEL HEPG2

• Main Author: Nia Arin Prasetyo, Vincentia
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/54780
• Institution: Institut Teknologi Bandung

Hepatitis B virus (HBV) belongs to Hepadnaviridae family and caused Hepatitis B infection. HBV infection can develop into liver carcinoma, one of which is due to presence of HBV X protein (HBx). Some mutations are known to be associated with development of liver carcinoma, for instance HBx K130M/V131I. In Indonesia, HBx T118N has been found at similar frequency as the HBx K130M/V131I. In previous studies, HBx T118N and K130M / V131I decreased the colony formation of HepG2. This study aims to examine the effect of T118N mutant HBx on B-cell lymphoma 2 (Bcl-2) expression in HepG2 cells using quantitative polymerase chain reaction (qPCR). First, Bcl-2 primer pair was designed, then analysed in silico using PRaTo and yielded value of -3. The primer specificity test showed single peak in qPCR melt curve. The qPCR efficiency measurement showed an efficiency value of 2.072. The plasmids containing gene encoding for wild type and mutant HBx were isolated and confirmed by migration analysis, restriction digest analysis, and nucleotide sequencing. HepG2 cells were transfected with the plasmids for 48 hours and the total RNA was isolated. The total RNA was reverse-transcribed into cDNA and used as a qPCR template. The level of Bcl-2 expression were determined using the relative quantification method Pfaffl. The presence of HBx T118N decreased Bcl-2 expression and the change was in line with the effect of HBx K130M/V131I.