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Hepatitis B virus (HBV) causes Hepatitis B and belongs to the family of Hepadnaviridae. Hepatitis B can develop into liver carcinoma through several mechanisms, one of which is mediated by the wild-type HBV X protein (HBx) and mutant HBx. Several HBx mutant are known to have a role in the developmen...
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| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/21802 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Hepatitis B virus (HBV) causes Hepatitis B and belongs to the family of Hepadnaviridae. Hepatitis B can develop into liver carcinoma through several mechanisms, one of which is mediated by the wild-type HBV X protein (HBx) and mutant HBx. Several HBx mutant are known to have a role in the development of liver carcinoma, i.e. HBx V5M/L, P38S, H94Y, I127T/N, and K130M/V131I. In Indonesia, HBx T118N is widely found in Hepatitis B patients but no further studies have been performed on the effect of HBx T118N on liver carcinoma. This study aims to determine the effect of HBx T118N on Bax expression in HepG2 cells and the result is used as the basis of counseling suggestions. First, Bax primer pair was designed using Primer3Plus, then analyzed in silico using PRaTo and showed value -5. The specificity of Bax primer was determined using quantitative polymerase chain reaction (qPCR), as well as its efficiency. The result showed that the designed primer was specific and the qPCR efficiency was 100,61%. The plasmids containing gene encoding for wild-type and mutant HBx were isolated, confirmed, and transfected to HepG2 cells for 48 hours. Total RNA was extracted from HepG2 cells, followed by DNase treatment and reverse transcription to synthesize cDNA. cDNA was then used as template to determine Bax expression. Bax expression analysis was performed using relative quantification method Pfaffl with GAPDH as normalisator. The result of this study showed that HBx T118N decreased Bax expression on HepG2 cell as compared to wild-type HBx, which was similiar to HBx K130M/V131I. This information serves as the basis of counseling suggestions given to health professional workers for early detection of hepatocellular carcinoma development. |
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