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Abstract

Beta-Amyrin Modulates P38 MAPK and Jnk Pathway to Inhibit Cell Proliferation and Induce ROS-mediated Apoptosis in HeLa Cells

Author(s): J. Anburaj1, Tamilselvi2*, Sanni Swapna3, Kamati Amuthavalli4
Department of Biotechnology, Karpagam Academy of Higher Education, Pollachi Main Road, Eachanari Post, Coimbatore, India, 1College of Horticulture, Tea Research Institute, Nanjing Agricultural University, Nanjing, Jiangsu, 2Institute and Department of Dermatology, The First Affiliated Hospital, Anhui Medical University, Hefei, Anhui, PR China, 3Department of Laboratory Services, Hindu Mission Hospital, Tambaram, Tamil Nadu, 4VRR Institute of Biomedical Science, Chennai, India

Correspondence Address:
Institute and Department of Dermatology, The First Affiliated Hospital, Anhui Medical University, Hefei, Anhui, PR China, E-mail: [email protected]

It is aimed to investigate the effect of β-amyrin on p38 mitogen-activated protein kinase and Jun N-terminal kinase pathways and apoptosis in HeLa cells. HeLa treated cells were divided into 6 groups, group I- HeLa untreated cells as control, group II- dimethyl sulfoxide serve as vehicle control, group III- cisplatin as standard drug, group IV- β-amyrin-treated HeLa cells, group V- cells were pretreated with 100 μm N-acetyl-L-cystein for 1 h and then treated with cisplatin and group VI- cells were pretreated with 100 μm N-acetyl-L-cystein for 1 h and then treated with β-amyrin. The antiproliferative effect was measured using the MTT assay. Genotoxic effects were studied using micronucleus assay. Total reactive oxygen species, nitric oxide and caspase 3 level were determined on a spectrofluorimeter and colorimeter. Protein expression was analyzed by immunoblotting. β-amyrin (10-200 μm) and cisplatin (0.01-100 μm) had an inhibitory effect on the proliferation of cancer cells in a dose-dependent manner, with the IC50 values at 100 μm and 10 μm for β-amyrin and cisplatin, respectively. Western blot analysis revealed expressions of apoptotic pathway related proteins, Bcl-2, caspase-3, caspase-9, phospho-p38 mitogen-activated protein kinase and phospho- Jun N-terminal kinase, growth arrest and deoxyribonucleic acid-damage-inducible, beta in all groups. Genotoxic effects were observed after treatment with β-amyrin as well as with cisplatin. It was observed that HeLa cells showed significant elevation of total reactive oxygen species after β-amyrin treatment. Protein expression analysis showed that the β-Amyrin upregulated phospho-p38 mitogen-activated protein kinase, phospho-Jun N-terminal kinase and growth arrest and deoxyribonucleic acid-damage-inducible, beta on HeLa cells. Increased phospho-Jun N-terminal kinase directly activated caspases and decreased Bcl-2 in HeLa cells. These results indicated that β-amyrin induced the apoptosis through reactive oxygen species-mediated mechanism by activating p38 mitogen-activated protein kinase and Jun N-terminal kinase through transcriptional factor, growth arrest and deoxyribonucleic acid-damage-inducible, beta. In turn, activated Jun N-terminal kinase directly activated caspase-9 and caspase-3 and destined the HeLa cells to apoptosis.

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