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All Journal HAYATI Journal of Biosciences Microbiology Indonesia Jurnal Matematika & Sains Pharmauho: Jurnal Farmasi, Sains, dan Kesehatan The Indonesian Biomedical Journal Jurnal Farmasi Indonesia YARSI Medical Journal
DEBBIE SOEFIE RETNONINGRUM
School of Pharmacy, Institut Teknologi Bandung
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Immunology & microbiology Mathematics Medicine & Pharmacology Public Health

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T118N Substitution of Hepatitis B X Protein Reduces Colony Formation of HepG2 Cells Artarini, Anita; Nurmalasari, Dewi Riskha; Permanasari, Silmi Citra; Riani, Catur; Tjandrawinata, Raymond Rubianto; Retnoningrum, Debbie Soefie
The Indonesian Biomedical Journal Vol 15, No 1 (2023)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v15i1.2095

Abstract

BACKGROUND: The acute Hepatitis B virus (HBV) infection usually ceases before six months, but chronic infection that lasts for more than six months might develop into liver cirrhosis and hepatocellular carcinoma (HCC). Viral particle load, HBV genotypes and association to the HBV x (HBx) gene mutations are the probable factors related to HCC occurrence. The mutation which leads to HBx T118N was found as the second most common HBx mutation in Indonesia, as compared to the known cancer-related HBx K130M/V131I mutant. However, the effect of T118N mutation and its combination with K130M/V131I on human hepatoma cells has not been elucidated well. Hence, this study was conducted to dissect the role of HBx T118N and its mutant combination in colony formation, as compared to the wild type HBx and cancer-related HBx K130M/V131I.METHODS: In this study, the genes encoding wild type HBx, HBx T118N, and HBx K130M/V131I mutations were obtained as synthetic gene. Meanwhile, the gene encoding HBx T118N/K130M/V131I mutations was successfully generated using site-directed mutagenesis. The optimum condition for colony formation assays was determined through Zeocin sensitivity test of HepG2 cells.RESULTS: Selection of HepG2 cells using Zeocin was determined at 200 µg/mL. Colony formation assays performed upon expression of HBx T118N and HBx T118N/K130M/V131I mutant proteins showed reduced colony numbers as compared to the expression of wild type HBx, similar to the effect from HBx K130M/V131I mutant expression.CONCLUSION: The HBx T118N and HBx T118N/K130M/V131I mutation caused less colony formation of HepG2 cells, similar to the K130/M131I mutation. This indicates a possible role of the T118N mutation in liver cancer development.KEYWORDS: colony formation assay, hepatitis B virus, HBx, T118N, K130M/V131I
Co-Authors ADAM REZA GANJARA Ana Indrayati ARTARINI, ANITA CATUR RIANI CHANDRA JINATA Chatib Warsa, Usman Dessy Natalia Dewi Riskha Nurmalasari Elfahmi Elfahmi, Elfahmi Elvi Restiawaty Endang Purwantini Enny Ratnaningsih ERNAWATI ARIFIN GIRI-RACHMAN Ernawati Arifin Giri-Rachman Fatmawaty Badaruddin Imam Supardi LARAS AJENG PITAYU Muhaimin Muhaimin Neni Nurainy Novia Syari Intan Oei Ban Liang Patricia Gita Naully Permanasari, Silmi Citra Putri Ayu Fajar Raymond R. Tjandrawinata ROGA FLORIDA KEMBAREN RUKMAN HERTADI Sukmadaja Asyarie Sumirtapura, Yeyet Cahyati Tan, Marselina Irasonia Tina Rostinawati Tri Suciati VALENTINA YURINA VALENTINA YURINA Wardono Niloperbowo Wojciechowska, Gladys Emmanuella Putri