Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5

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_version_ 1860799433213476864
building INTELEK Repository
collection Online Access
collectionurl https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection407072
date 2015-06-24 09:33:37
eventvenue Johor Bahru, Malaysia
format Restricted Document
id 5986
institution UniSZA
originalfilename 0718-01-FH03-FBIM-15-03281.pdf
person Hadi Nur
recordtype oai_dc
resourceurl https://intelek.unisza.edu.my/intelek/pages/view.php?ref=5986
spelling 5986 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=5986 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection407072 Restricted Document Conference Conference Paper application/pdf 6 1.6 Adobe Acrobat Pro DC 20 Paper Capture Plug-in Hadi Nur 2015-06-24 09:33:37 0718-01-FH03-FBIM-15-03281.pdf UniSZA Private Access Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5 Antimicrobial peptides (AMPs) have been reported to display multifunctional properties as potential therapeutic agents. They exhibit rapid killing and a broad spectrum of activity against Gram-positive and Gram-negative bacteria, fungi, parasites, enveloped viruses and tumour cells. An AMP, hepcidin TH1-5 (HepTH1-5) is a small and cationic peptide was shown as important innate immune defense in Tilapia (Oreochromis mossambicus). Recent studies have shown that synthetically derived HepTH1-5 displayed cytotoxic effect on tested cancer cell lines. In this study, we aimed to investigate the cytotoxic and apoptosis effects of HepTH1-5 on HL-60, HepG2 and HeLa cells and compare to noncancerous cell lines, Chang and NIH/3T3. We also investigated the effect of this peptide on cell cycle. Cytotoxic assay by MTT revealed that HepTH1-5 is highly cytotoxic on HepG2, HeLa and HL-60 cells with IC50 of 8.58 x10-4 µM, 0.011 µM and 0.0858 µM respectively. These results also comparable with methotrexate, a standard chemotherapy drug. Intriguingly, it does not show significant cytotoxicity against normal cells. Apoptogenic effects of HepTH1-5 were studied by acridine orange/propidium iodide staining and revealed that after 24h treatment, 61.2% of HepG2 cells were in early apoptosis, 20.7% in late apoptosis while 1% of cells were in necrosis. In contrast, after 24h treatment with methotrexate, only 18.2% of cells were in early apoptosis, whereas 76.3% were in late apoptosis. This trend was continued after 48h treatment but prolonged incubation of 72h saw a dramatic change in the apoptotic effect where less than 12% of cells were at early apoptosis while more than 79.7% were at late apoptosis. Cell cycle analysis on HepG2 cells showed that HepTH1-5 was able to block cell at G2/M phase significantly but not in HeLa while methotrexate arrest cells at G0/G1 phase. This data revealed that apoptosis effect of HepTH1-5 and methotrexate in cell cycle might occur at different mechanism. Taking all the data together, it was found that HepTH1-5 significantly triggers apoptosis in cancerous cells and it shows that this peptide could potentially be developed as anticancer agent. International Conference on Natural Product 2015 Johor Bahru, Malaysia
spellingShingle Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
summary Antimicrobial peptides (AMPs) have been reported to display multifunctional properties as potential therapeutic agents. They exhibit rapid killing and a broad spectrum of activity against Gram-positive and Gram-negative bacteria, fungi, parasites, enveloped viruses and tumour cells. An AMP, hepcidin TH1-5 (HepTH1-5) is a small and cationic peptide was shown as important innate immune defense in Tilapia (Oreochromis mossambicus). Recent studies have shown that synthetically derived HepTH1-5 displayed cytotoxic effect on tested cancer cell lines. In this study, we aimed to investigate the cytotoxic and apoptosis effects of HepTH1-5 on HL-60, HepG2 and HeLa cells and compare to noncancerous cell lines, Chang and NIH/3T3. We also investigated the effect of this peptide on cell cycle. Cytotoxic assay by MTT revealed that HepTH1-5 is highly cytotoxic on HepG2, HeLa and HL-60 cells with IC50 of 8.58 x10-4 µM, 0.011 µM and 0.0858 µM respectively. These results also comparable with methotrexate, a standard chemotherapy drug. Intriguingly, it does not show significant cytotoxicity against normal cells. Apoptogenic effects of HepTH1-5 were studied by acridine orange/propidium iodide staining and revealed that after 24h treatment, 61.2% of HepG2 cells were in early apoptosis, 20.7% in late apoptosis while 1% of cells were in necrosis. In contrast, after 24h treatment with methotrexate, only 18.2% of cells were in early apoptosis, whereas 76.3% were in late apoptosis. This trend was continued after 48h treatment but prolonged incubation of 72h saw a dramatic change in the apoptotic effect where less than 12% of cells were at early apoptosis while more than 79.7% were at late apoptosis. Cell cycle analysis on HepG2 cells showed that HepTH1-5 was able to block cell at G2/M phase significantly but not in HeLa while methotrexate arrest cells at G0/G1 phase. This data revealed that apoptosis effect of HepTH1-5 and methotrexate in cell cycle might occur at different mechanism. Taking all the data together, it was found that HepTH1-5 significantly triggers apoptosis in cancerous cells and it shows that this peptide could potentially be developed as anticancer agent.
title Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
title_full Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
title_fullStr Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
title_full_unstemmed Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
title_short Induction of Apoptosis and G2/M Phase Cell Cycle Arrest by Antimicrobial Peptide HepTH1-5
title_sort induction of apoptosis and g2/m phase cell cycle arrest by antimicrobial peptide hepth1-5