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Journal : The Indonesian Biomedical Journal

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 Achmad K. Harzif Agung Endro Nugroho Agung Endro Nugroho Andon Hestiantoro Andri Prasetiyo Anindini Winda Amalia Anindyajati Anindyajati Anita Artarini Armini Syamsidi ARTARINI, ANITA B. Atawolo, Andreas Babu, Murali Mohan Budi Wiweko Caroline G. Puspita CATUR RIANI Danang Agung Yunaidi Dayana, Hepy Debbie S. Retnoningrum Debbie S. Retroningrum DEBBIE SOEFIE RETNONINGRUM Dewi Riskha Nurmalasari Diana Nur Afifah Esti Mumpuni Esti Mumpuni Esti Mumpuni, Esti Gayatri, Anggi Gita Pratama Hapsari, Riani Hartati Tuna, Hartati Heliany, Ina Henry Soelistyo Budi Henry Soelistyo Budi, Henry Soelistyo Herbert Situmorang Herbert Situmorang Huda Shalahudin Darusman Indriawati, Iin Irsan Hasan Ismaya, Wangsa T. Kaka Renaldi Kanadi Sumapradja Krisnayanti, Ni Putu Eka Kristiana, Hery Kristiana, Hery Laurentia Stephani Liana W. Susanto Lilik Sulastri Luh Putu Nurshanti, Ni Maggy T. Suhartono Maggy T. Suhartono Maggy Thenawidjaja Suhartono Marcellus Simadibrata Martha Wijaya, Harry Melva Louisa Murdani Abdullah Nafrialdi Nafrialdi Nailufar, Florensia Nailufar, Florensia Novia Tri Astuti Novia Tri Astuti Novitasari, Putri Rachma Permanasari, Silmi Citra Rachman, Andika Raden Muharam Natadisastra Retroningrum, Debbie S. Santi Tan Saut Horas Hatoguan Nababan Setiabudy, Rahajuningsih D Shirly Kumala SILMI MARIYA Simatupang, Stefeny Theresia Sinaga, Wenny S.L. Soelistyo Budi, Henry Susanto, Liana W Suwijiyo Pramono Suwijiyo Pramono Tan, Santi Viedya Novalinda Said Wangsa T. Ismaya Wangsa Tirta Ismaya Widjaja, Nadia Winoto, Imelda L Winoto, Imelda L Yanti Lim Zaelani, Bella Fatima Dora