A study of the potential anticancer activity of Mangifera zeylanica bark: Evaluation of cytotoxic and apoptotic effects of the hexane extract and bioassay-guided fractionation to identify phytochemical constituents

Ediriweera,Meran  Keshawa; Tennekoon,Kamani  Hemamala; Samarakoon,Sameera  Ranganath; Thabrew,Ira; Dilip De Silva,Egodage

doi:10.3892/ol.2016.4087

February-2016 Volume 11 Issue 2

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February-2016 Volume 11 Issue 2

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Article Open Access

A study of the potential anticancer activity of Mangifera zeylanica bark: Evaluation of cytotoxic and apoptotic effects of the hexane extract and bioassay-guided fractionation to identify phytochemical constituents

Authors:
- Meran Keshawa Ediriweera
- Kamani Hemamala Tennekoon
- Sameera Ranganath Samarakoon
- Ira Thabrew
- Egodage Dilip De Silva
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Affiliations: Institute of Biochemistry, Molecular Biology and Biotechnology, University of Colombo, Colombo 00300, Sri Lanka, Department of Chemistry, Faculty of Science, University of Colombo, Colombo 00300, Sri Lanka

Copyright: © Ediriweera et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 1335-1344
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Published online on: January 8, 2016

https://doi.org/10.3892/ol.2016.4087
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Abstract

The present study investigated the potential anticancer activity of the bark of Mangifera zeylanica, an endemic plant in Sri Lanka that has been traditionally used for cancer therapy. Cytotoxic and apoptotic effects were investigated in vitro using sulphorodamine assay, acridine orange and ethidium bromide staining, caspase‑3 and ‑7 activity, DNA fragmentation and reverse transcription‑quantitative polymerase chain reaction in estrogen receptor positive MCF‑7 and triple‑negative MDA‑MB‑231 breast cancer cell lines, SKOV‑3 ovarian cancer cell line and MCF‑10A normal mammary epithelial cells. Hexane extract demonstrated increased levels of cytotoxicity in cancer cells (IC50, 86.6‑116.5 µg/ml) compared with normal cells (IC50, 217.2 µg/ml). Chloroform extract demonstrated increased cytotoxicity to normal cells (IC50, 92.9 µg/ml) compared with cancer cells (IC50, 280.1‑506.5 µg/ml). Exposure to the hexane extract led to morphological changes characteristic of apoptosis and DNA fragmentation in the three cancer cell lines. Caspase‑3 and ‑7 were significantly activated in MDA‑MB‑231 and SKOV‑3 cells, indicating the occurrence of caspase‑dependent apoptosis in these cells, and caspase‑independent apoptosis in MCF‑7 cells. Furthermore, upregulation of proapoptotic Bcl‑2‑associated X protein occurred in the three cancer cell lines, and antiapoptotic survivin was downregulated in MCF‑7 and SKOV‑3 cells; by contrast, tumor protein p53 was upregulated only in MCF‑7 cells, suggesting p53‑mediated apoptosis in MCF‑7 cells and p53‑independent apoptosis in the remaining cancerous cell lines. In addition, fraction M1 obtained from bioactivity‑guided fractionation of the hexane extract demonstrated increased cytotoxicity in cancer cells (IC50, 15.4‑38.7 µg/ml) compared with normal cells (IC50, 114.6 µg/ml), with the highest cytotoxicity observed in MDA‑MB‑231 triple‑negative breast cancer cells. The hexane extract of M. zeylanica bark contained polyphenols and flavonoids, and caused free radical scavenging activity. Its gas chromatography‑mass spectrometry profile revealed the presence of long‑chain hydrocarbons, including β‑sitosterol and β‑amyrin. Fraction M1 contained seven unknown compounds and a small number of known non‑cytotoxic compounds. Collectively, results obtained in the present study indicate that the hexane extract of M. zeylanica bark mediates cytotoxic activities through induction of apoptosis in three cancer cell lines; thus, the hexane extract may be used to isolate novel anti-cancer compounds.

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