Antiproliferative and proapoptotic activities of marine sponge Hyrtios erectus extract on breast carcinoma cell line (MCF-7)
Ramachandran Muthiyan1, Balwin Nambikkairaj1, Nilkamal Mahanta2, Titus Immanuel3, Rahul Shubhra Mandal4, Kubendiran Kumaran5, Arun Kumar De6
1 Department of Zoology, Voorhees College, Thiruvalluvar University, Vellore, India
2 Department of Chemistry, Institute for Genomic Biology, University of Illinois, Urbana-Champaign, Illinois, USA
3 Division of Fisheries Sciences, Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India
4 Biomedical Informatics Centre, National Institute of Cholera and Enteric Diseases, Beliaghata, Kolkata, India
5 Syngene International Limited (Biocon), Biocon Park, Bangalore, India
6 Department of Animal Sciences, Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India; Department of Animal Sciences, University of Illinois, Urbana-Champaign, Illinois, USA
Dr. Arun Kumar De
Department of Animal Sciences,Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, India
Source of Support: None, Conflict of Interest: None
Background: Marine sponge is a rich natural resource of many pharmacologically important compounds. Objective: Marine sponge Hyrtios erectus, collected from North Bay, South Andaman Sea, India, was screened for potential antiproliferative and proapoptotic properties on a breast adenocarcinoma cell line (MCF-7). Materials and Methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to test the antiproliferative and cytotoxicity effects of the sponge extract. Analysis of apoptosis and cell cycle stages were done by flow cytometry. The expression of several apoptotic-related proteins in MCF-7 cells treated by the extract was evaluated by Western blot analysis. Various analytical techniques including Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry, and nuclear magnetic resonance were employed to determine the identity of the active compounds in the sponge extract. Results: N-Hexane extract of the sponge inhibited proliferation of the MCF-7 cell line in a dose- and time-dependent manner. Exposure of the sponge extract triggered apoptosis of the MCF-7 cells, induced DNA fragmentation, and arrested the cells in G2/M phase. Treatment of the sponge extract induced downregulation of antiapoptotic Bcl-2 protein and upregulation of Bax, caspase-3, caspase-9, and fragmented poly(ADP ribose)polymerase proteins in MCF-7 cells. Five bioactive compounds have been identified in the extract. Conclusion: The antiproliferative and proapoptotic activities of the tested extract suggested the pharmacologic potential of the identified compounds. Further characterization of the identified compounds are in progress.
Abbreviations used: GC-MS: Gas chromatography-mass spectrometry; FT-IR: Fourier transform infrared spectroscopy; NMR: Nuclear magnetic resonance; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.