INDUCTION OF CASPASE-3 DEPENDENT APOPTOSIS, CELL CYCLE ARREST AND CYTOTOXICITY IN BREAST CANCER CELLS BY ABRUS PRECATORIUS
DOI:
https://doi.org/10.22159/ijpps.2018v10i8.17996Keywords:
Abrus precatorius L, Cytotoxicity, Breast cancer cells, ApoptosisAbstract
Objective: To evaluate the leaf extract of Abrus precatorius as a potential therapy for breast cancer treatment.
Methods: Aqueous leaf extracts of A. precatorius was prepared by the process of maceration. The collected filtrate was further partitioned successively into five solvent fractions starting from water, hexane, chloroform, ethyl acetate and butanol by solvent fractionation method using separating funnel. Finally, all the five fractions were subjected to cytotoxic activity assay. The molecular mechanism underlying cytotoxicity was determined by using various approaches viz., 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Fluorescence-activated cell sorting (FACS) analysis, Western blot analysis, casp-glow assay and Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis.
Results: Among five fractions only ethyl acetate fraction of A. precatorius (EAF-AP) showed significant cytotoxic activity on MDA-MB-231 cells, with an IC50 value of 47.3 µg/ml. Apoptosis was confirmed by the appearance of Sub G0/G1 (apoptotic) peak by FACS analysis. Western blotting results clearly indicated cleavage of Caspase-3 and PARP. Casp-glow assay confirmed activation of caspase-3, an important mediator of apoptosis. Semi-quantitative RT-PCR analysis showed an up-regulation of pro-apoptotic genes (p21, p53 and Bax) and down-regulation of the anti-apoptotic Bcl-2 gene, which is an important hallmark of an apoptosis.
Conclusion: All these results indicate that EAF-AP, induced apoptosis in MDA-MB-231 cells, making A. precatorius a potentially good candidate for anticancer drug development.
Downloads
References
Weinberg RA. Oncogenes and the molecular biology of cancer. J Cell Biol 1983;97:1661-2.
Bertram JS. The molecular biology of cancer. Mol Aspects Med 2001;21:167-223.
Grimm M, Cetindis M, Lehmann M, Biegner T, Munz A, Teriete P, et al. Association of cancer metabolism-related proteins with oral carcinogenesis–indications for chemoprevention and metabolic sensitizing of oral squamous cell carcinoma. J Transl Med 2014;12:208.
Globocan. Estimated cancer incidence, mortality and prevalence worldwide in International Agency for Research on Cancer (IARC); 2012.
World Health Organization. World Cancer Report. The International agency for research on cancer (IARC), Geneva; 2014.
Ahmedin J, Rebeca S, Elizabeth W, Yongping H, Xu J, Michael JT. Cancer statistics. CA Cancer J Clin 2009;59:225-49.
World Health Organization. Cancer incidence and mortality worldwide. The International agency for research on cancer (IARC), Geneva; 2008.
Brody JG, Rudel RA. Environmental pollutants and breast cancer. Environ Health Persp 2003;111:1007–19.
Moorthi C, Kumar CS, Kathiresan K. Synergistic anti-cancer activity of curcumin and bio-enhancers combination against various cancer cell lines. Int J Pharm Pharm Sci 2014;6:901-3.
Altmann KH, Gertsch J. Anticancer drugs from nature-natural products as a unique source of new microtubule-stabilizing agents. Nat Prod Rep 2007;24:327-57.
Surekha R, Deshpande SNB. Cytotoxicity of stem extracts of selected cassia species against Hela and breast cancer cell lines in vitro. Asian J Pharm Clin Res 2017;10:80-2.
Cragg GM, Newman J. Nature: a vital source of leads for anticancer drug development. Phytochem Rev 2009;8:313–31.
Upur H, Yusup A, Baudrimont I, Umar A, Berke B, Yimit D, et al. Inhibition of cell growth and cellular protein, DNA and RNA synthesis in human hepatoma (HepG2) cells by ethanol extract of abnormal Savda Munziq of traditional Uighur medicine. Evid Based Complement Alternat Med 2011:1-9. http://dx.doi.org/ 10.1093/ecam/nen062
Nadkarni KM. Indian Materia Medica. Popular Prakashan Bombay, India; 1976.
Adedapo AA, Omoloye OA, Ohore OG. Studies on the toxicity of an aqueous extract of the leaves of A. precatorius in rats. Onderstepoort J Vet Res 2007;74:31–6.
Dnyaneshwar JT, Ravindra YP. Effect of A. precatorius leaves on milk induced leukocytosis and eosinophilia in the management of asthma. Asian Pac J Trop Med 2012;1:40-2.
Shih SF, Wu YH, Hung CH, Yang HY, Lin JY. Abrin triggers cell death by inactivating a thiol-specific antioxidant protein. J Biol Chem 2001;276:21870–7.
Narayanan S, Surolia A, Karande AA. Ribosome-inactivating protein and apoptosis: abrin causes cell death via the mitochondrial pathway in Jurkat cells. Biochem J 2004;377:233–40.
Bhutia SK, Mallick SK, Maiti TK. In vitro immunostimulatory properties of Abrus lectins derived peptides in tumor-bearing mice. Phytomedicine 2009;16:776–82.
Mojarrab M, Lagzianb MS, Emamic SA, Asilic J, Najaranb ZT. In vitro antiproliferative and apoptotic activity of different fractions of Artemisia armeniaca. Rev Bras Farmacogn 2013;23:783-8.
Mosmann T. Rapid colourimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;16:55–63.
Plumb JA. Cell sensitivity assays: the MTT assay. Methods Mol Med 2004;88:165-9.
Schwartsmann G. Marine organisms and other novel natural sources of new cancer drugs. Ann Oncol 2000;11:235-43.
Lall RK, Syed DN, Adhami VM, Khan MI, Mukhtar H. Dietary polyphenols in the prevention and treatment of prostate cancer. Int J Mol Sci 2015;16:3350-76.
Polachi N, Nagaraja P, Subramaniyan B, Mathan G. Antiproliferative activity of n-butanol floral extract from Butea monosperma against Hct 116 colon cancer cells; drug-likeness properties and in silico evaluation of their active compounds toward glycogen synthase kinase-3β/axin and β-catenin/t-cell factor-4 protein complex. Asian J Pharm Clin Res 2015;8:134–41.
Christopher SP. The significance of spontaneous and induced apoptosis in the gastrointestinal tract of mice. Cancer Metast Rev 1992;11:179-95.
Antony E, Sathiavelu M, Arunachalam S. Synthesis of silver nanoparticles from the medicinal plant Bauhinia acuminata and Biophytum sensitivum–a comparative study of its biological activities with plant extract. Int J Appl Pharm 2017;9:22-9.
Dobashi Y, Takehana T, Ooi A. Perspectives on cancer therapy: cell cycle blockers and perturbators. Curr Med Chem 2003;10:2549-58.
Zeytinoglu H, Incesu Z, Baser KH. Inhibition of DNA synthesis by carvacrol in mouse myoblast cells bearing a human N-RAS oncogene. Phytomedicine 2003;10:292-9.
Kim DI, Lee SJ, Lee SB, Park K, Kim WJ, Moon SK. The requirement for Ras/Raf/ERK pathway in naringin induced G1 cell cycle arrest via p21WAF1 expression. Carcinogenesis 2008;29:1701-9.
Kanzawa T, Germano IM, Komata T, Ito H, Kondo Y, Kondo S. Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ 2004;11:448-57.
Kim H, Yong JM, Seok AK, Cho SK. Quercetin induces mitochondrial-mediated apoptosis and protective autophagy in human glioblastoma U373MG cells. Oxid Med Cell Longev 2013;596496:1-10.
Soldani C, Scovassi AI. Poly (ADP-ribose) polymerase-1 cleavage during apoptosis: an update. Apoptosis 2002;7:321-8.
Jin CY, Park C, Cheong J, Choi BT, Lee TH, Lee JD, et al. Genistein sensitizes TRAIL-resistant human gastric adenocarcinoma AGS cells through activation of caspase-3. Cancer Lett 2007;257:56-64.
Daniel RC, Bernand WS. Etoposide-induced cytotoxicity in two human T-cell leukaemic lines: delayed loss of membrane permeability rather than DNA fragmentation as an indicator of programmed cell death. Cancer Res 1993;53:4887-96.
Vogelstein B, Kinzler KW. p53 function and dysfunction. Cell 1992;70:523-6.
Korsmeyer SJ, Shutter JR, Veis DJ, Merry DE, Oltvai ZN. Bcl-2/Bax: a rheostat that regulates an antioxidant pathway and cell death. Sem Cancer Biol 1993;4:327-32.
Aggarwal F, Conforti G, Ioele GA, Statti M, Marrelli Ragno G, Menichini F. Antiproliferative activity against human tumor cell lines and toxicity test on Mediterranean dietary plants. Food Chem Toxicol 2008;46:3325-32.
Published
How to Cite
Issue
Section
