LAWSONIA ALBA LEAVES INDUCE APOPTOSIS AND CELL CYCLE ARREST IN B16F10 MELANOMA CELLS
DOI:
https://doi.org/10.22159/ijpps.2018v10i5.24526Keywords:
Lawsonia alba, Cancer, Melanoma, ApoptosisAbstract
Objective: The present study was designed to investigate the anti-melanoma activity of the ethyl acetate fraction of Lawsonia alba lam leaves (ELA) against B16F10 cells.
Methods: Cytotoxicity of ELA on B16F10 cells was determined by MTT assay and supported with the morphology of apoptotic and necrotic cells under phase-contrast microscope, fluorescence microscopy with AO/EtBr, confocal microscopy with PI, Agarose gel electrophoresis and Annexin V-FITC/PI staining, mitochondrial membrane potential and cell cycle arrest by FACS was also performed on B16F10 cells.
Results: Cytotoxic effect of ELA on B16F10 melanoma cell was confirmed by MTT assay with an IC50 value of 14.10μg/ml. Morphological study showed arrays of both the nuclear changes including chromatin condensation and apoptotic body formation indicating that the treatment with ELA and 5-Fluorouracil (standard) causes apoptotic changes in the melanoma cells compared to the untreated control. Agarose gel electrophoresis study showed fragmented DNA in the form of ladder. The depolarization of mitochondrial membrane potential was confirmed. Flow cytometric analysis showed appreciable number of cells in early apoptotic stage. The cells were arrested mostly in G0/G1 phase of cell cycle.
Conclusion: Ethyl acetate fraction of Lawsonia alba L. leaves possesses potent apoptotic activity against B16F10 cells.
Downloads
References
Markovic SN, Erickson LA, Rao RD. Melanoma study group of the mayo clinic cancer center. Malignant melanoma in the 21st century, Part 1: epidemiology, risk factors, screening, prevention, and diagnosis. Mayo Clin Proc 2007;82:364–80.
Vikey AK, Vikey D. Primary malignant melanoma, of head and neck: a comprehensive review of the literature. Oral Oncol 2012;48:399–403.
Guevara Canales JO, Gutierrez Morales MM, Sacsaquispe Contreras SJ, Sanchez Lihon J, Morales Vadillo R. Malignant melanoma of the oral cavity. Review of the literature and experience in a peruvian population. Med Oral Patol Oral Cir Bucal 2012;17:206–11.
Bhatia S, Tykodi SS, Thompson JA. Treatment of metastatic melanoma: an overview. Oncology 2009;23:488–96.
Grossman D, Altieri DC. Drug resistance in melanoma: mechanisms, apoptosis, and new potential therapeutic targets. Cancer Metastasis Rev 2001;20:3-11.
Hassan M, Watari H, AbuAlmaaty A, Ohba Y, Sakuragi N. Apoptosis and molecular targeting therapy in cancer. Biomed Res Int 2014. http://dx.doi.org/10.1155/2014/150845
Arun P, Purushotham KG, Jayarani JJ, D Kumari V. In vitro antibacterial activity and flavonoid contents of Lawsonia inermis (Henna). Int J PharmTech Res 2010;2:1178-81.
Babili FE, Valentin A, Chatelain C. Lawsonia inermis: its anatomy and its antimalarial, antioxidant and human breast cancer cells MCF7 activities. Pharm Anal Acta 2013;4:203.
Dutta S, Deb N, Pattanaik AK, Besra SE. The apoptosis-inducing potential of Lawsonia alba lam. Leaves on hepatocellular carcinoma (Hep-G2) cells along with its anti-oxidant property. Int J Pharm Pharm Sci 2016;8:156-62.
Kapadia GJ, Rao GS, Sridhar R, Ichiishi E, Takasaki M, Suzuki N, et al. Chemoprevention of skin cancer: effect of Lawsonia inermis L. (Henna) leaf powder and its pigment artifact, lawsone in the epstein-barr virus early antigen activation assay and in two-stage mouse skin carcinogenesis models. Adv Anticancer Agents Med Chem 2013;13:1500-7.
Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 1986;89:271-7.
Fontes JEN, Ferraz RP, Britto AC, Carvalho AA, Moraes MO, Pessoa C, et al. Antitumor effect of the essential oil from leaves of Guatteria pogonopus (Annonaceae). Chem Biodivers 2013;10:722–9.
Jimenez PC, Wilke DV, Takeara R, Lotufo TM, Pessoa C. Cytotoxicity activity of a dichloromethane fraction and fractions obtained from Eudistoma vannamei (Tunicata: Ascidiacea). Comp Biochem Physiol MolIntegr Physiol 2008;151:391-8.
Gupta SD, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni JR, et al. Indian black scorpion (Heterometrus bengalensis Koch) venom-induced anti-proliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leuk Res 2007;31:817–25.
Walker PR, Kokileva L, LeBlanc J, Sikorska M. Detection of the initial stages of DNA fragmentation in apoptosis. BioTechniques 1993;15:1032-40.
Liao PC, Lieu PC. Cell cycle specific induction of apoptosis and necrosis by paclitaxel in the leukemic U937 cells. Life Sci 2005;76:1623-39.
Deborah R, Nathaniel BG, David AN, Yuqun GS. A simple technique for quantifying apoptosis in 96 well plates. BMC Biotechnol 2005;5:12.
Telford WG, King LE, Fraker PJ. Induction of apoptosis has been a new target for innovative mechanism-based drug discovery. Cell Prolif 1991;24:447–59.
Vermes I, Haanen C, Steffens Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 1995;184:39-51.
Brauchle E, Thude S, Brucker SY, Schenke Layland K. Cell death stages in single apoptotic and necrotic cells monitored by Raman microspectroscopy. Sci Rep 2014;4:1-9.
Dartsch DC, Schaefer A, Boldt S, Kolch W, Marquardt H. Comparison of anthracyclines induced death of human leukemia cells: programmed cell death versus necrosis. Apoptosis 2002;7:537–48.
Published
How to Cite
Issue
Section
