Methanolic extract of Abrus precatorius promotes breast cancer MDA-MB-231 cell death by inducing cell cycle arrest at G_0/G_1 and upregulating Bax
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摘要
Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate and methanol by Soxhlet extraction.Aqueous extract was prepared by decoction at 50 ℃.Extracts of A.precatorius leaves were used to treat selected cancer and normal cell lines for72 h.Furthermore,3-(4,5-dimethyl thiazol-2-yl)2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability.Analysis of cell cycle arrest,apoptosis assay and apoptosis protein expressions were determined by flow cytometry.Results:Methanolic extract of A.precatorius leaves showed the lowest IC_(50) on MDA-MB-231 cells at(26.40±5.40)μg/mL.Flow cytometry analysis revealed that cell arrest occurred at G_0/G_1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB-231 cells treated with methanolic extract of A.precatorius leaves.Methanolic extract of A.precatorius leaves induced apoptosis by upregulation of Bax,p53 and caspase-3 and downregulation of Bcl-2.Conclusions:Methanolic extract of A precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway.
Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate and methanol by Soxhlet extraction.Aqueous extract was prepared by decoction at 50 ℃.Extracts of A.precatorius leaves were used to treat selected cancer and normal cell lines for72 h.Furthermore,3-(4,5-dimethyl thiazol-2-yl)2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability.Analysis of cell cycle arrest,apoptosis assay and apoptosis protein expressions were determined by flow cytometry.Results:Methanolic extract of A.precatorius leaves showed the lowest IC_(50) on MDA-MB-231 cells at(26.40±5.40)μg/mL.Flow cytometry analysis revealed that cell arrest occurred at G_0/G_1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB-231 cells treated with methanolic extract of A.precatorius leaves.Methanolic extract of A.precatorius leaves induced apoptosis by upregulation of Bax,p53 and caspase-3 and downregulation of Bcl-2.Conclusions:Methanolic extract of A precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway.
Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate and methanol by Soxhlet extraction.Aqueous extract was prepared by decoction at 50 ℃.Extracts of A.precatorius leaves were used to treat selected cancer and normal cell lines for72 h.Furthermore,3-(4,5-dimethyl thiazol-2-yl)2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability.Analysis of cell cycle arrest,apoptosis assay and apoptosis protein expressions were determined by flow cytometry.Results:Methanolic extract of A.precatorius leaves showed the lowest IC_(50) on MDA-MB-231 cells at(26.40±5.40)μg/mL.Flow cytometry analysis revealed that cell arrest occurred at G_0/G_1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB-231 cells treated with methanolic extract of A.precatorius leaves.Methanolic extract of A.precatorius leaves induced apoptosis by upregulation of Bax,p53 and caspase-3 and downregulation of Bcl-2.Conclusions:Methanolic extract of A precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway.
引文
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[14]Geran R,Greenberg N,MacDonald M,Schumacher A,Abbott256B.National Cancer Institute protocols for screening of anticancer compounds.Cancer Chemother Rep 1972;3:1-103.
[15]Igarashi M,Miyazawa T.The growth inhibitory effect of conjugated linoleic acid on a human hepatoma cell line,HepG2,is induced by a change in fatty acid metabolism,but not the facilitation of lipid peroxidation in the cells.Biochim Biophys Acta 2001;1530(2-3):162-171.
[16]Sharma A,Kaur R,Katnoria JK,Kaur R,Nagpal AK.Family Fabaceae:A boon for cancer therapy.In:Malik S(ed.)Biotechnology and production of anti-cancer compounds.Springer;2017,p.157-175.
[17]Rungruangmaitree R,Jiraungkoorskul W.Pea,Pisum sativum,and its anticancer activity.Phcog Rev 2017;11(21):39-42.
[18]Lee JH,Kim KS,Lee YC,Cho CW,Kim KM,Chung YS.Anticancer composition containing herbal extract.US 2011/0293754 A1(Patent)2017.
[19]Ahmad F,Anwar F,Hira S.Review on medicinal importance of fabaceae family.Ph OL 2016;3:151-156.
[20]Kim SJ,Min HY,Chung HJ,Park EJ,Hong JY,Kang YJ,et al.Inhibition of cell proliferation through cell cycle arrest and apoptosis by thio-ClIB-MECA,a novel A3 adenosine receptor agonist,in human lung cancer cells.Cancer Lett 2008;264(2):309-315.
[21]Li A,Blow JJ.The origin of CDK regulation.Nat Cell Biol 2001;3(8):E182-184.
[22]Czabotar PE,Lessene G,Strasser A,Adams JM.Control of apoptosis by the BCL-2 protein family:Implications for physiology and therapy.Nat Rev Mol Cell Biol 2014;15(1):49-63.
[23]Bai L,Wang S.Targeting apoptosis pathways for new cancer therapeutics.Annu Rev Med 2014;65:139-155.
[24]Chien SY,Wu YC,Chung JG,Yang JS,Lu HF,Tsao MF,et al.Quercetininduced apoptosis acts through mitochondrial-and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells.Hum Exp Toxicol2009;28(8):493-503.
[25]Pepper C,Hoy T,Bentley P.Elevated Bcl-2/Bax are a consistent feature of apoptosis resistance in B-cell chronic lymphocytic leukaemia and are correlated with in vivo chemoresistance.Leuk Lymphoma 1998;28(3-4):355-361.
[26]Beroukhim R,Mermei CH,Porter D,Wei G,Raychaudhuri S,Donovan J,et al.The landscape of somatic copy-number alteration across human cancers.Nature 2010;463(7283):899-905.
[27]Castle VP,Heidelberger KP,Bromberg J,Ou X,Dole M,Nu?ez G.Expression of the apoptosis-suppressing protein bcl-2,in neuroblastoma is associated with unfavorable histology and N-myc amplification.Am JPathol 1993;143(6):1543-1550.
[28]Olopade OI,Adeyanju MO,Safa AR,Hagos F,Mick R,Thompson CB,et al.Overexpression of BCL-x protein in primary breast cancer is associated with high tumor grade and nodal metastases.Cancer J Sci Am1997;3(4):230-237.
[29]Tait SW,Green DR.Mitochondria and cell death:Outer membrane permeabilization and beyond.Nature Rev Mol Cell Biol 2010;11(9):621-632.
[30]McIlwain DR,Berger T,Mak TW.Caspase functions in cell death and disease.Cold Spring Harb Perspect Biol 2013;5(4):a008656.Doi:10.1101/cshperspect.a008656.
[2]Cao K,Tait SW.Apoptosis and cancer:Force awakens,phantom menace,or both?.In:Galluzzi L(ed.)International review of cell and molecular biology.Elsevier;2018,p.135-152.
[3]Sreelatha SA,Jeyachitra,Padma PR.Antiproliferation and induction of apoptosis by Moringa oleifera leaf extract on human cancer cells.Food Chem Toxicol 2011;49(6):1270-1275.
[4]World Health Organization.Cancer.[Online]Available from:https://www.who.int/news-room/fact-sheets/detail/cancer.[Accessed on 5th February2019].
[5]World Health Organization.Cancer today:Data visualization tools for exploring the global cancer burden in 2018.[Online]Available from:https://gco.iarc.fr/today/home.[Accessed on 5th February 2019].
[6]Wan-Ibrahim WS,Ismail TNT,Mohd-Salleh SF,Ismail N.GC-MSanalysis of phytochemical compounds in aqueous leaf extract of Abrus precatorius.Pertanika J Trop Agric Sci 2018;41(1):241-250.
[7]Gul MZ,Manjulatha K,Bhat MY,Maurya R,Qureshi IA,Ghazi IA.Antiproliferative and apoptosis-inducing effects of Abrus precatorius against human monocytic leukaemia(THP-1)cell line.Indian J Pharm Sci 2018;80(2):307-317.
[8]Sofi MS,Sateesh MK,Bashir M,Ganie MA,Nabi S.Chemopreventive and anti-breast cancer activity of compounds isolated from leaves of Abrus precatorius L.3 Biotech 2018;8(8):371.
[9]Lebri M,Tilaoui M,Bahi C,Achibat H,Akhramez S,Fofie YBN,et al.Phytochemical analysis and in vitro anticancer effect of aqueous extract of Abrus precatorius Linn.Der Pharma Chemica 2015;7(8):112-117.
[10]Ghosh T,Mitra P,Jha DK,Mitra PK.Abrus precatorius linnaeus and its biological activities-a review.World J Pharm Pharm Sci 2017;6(8):352-368.
[11]Oladimeji AO,Babatunde O,Musa RT,M’civer FA,Lawal AT,Oguwande IA.GC-MS analysis and cytotoxic activity of essential oils from the leaves of Abrus precatorius L.Gaertn.Asian Pac J Trop Dis 2016;6(5):372-375.
[12]Gul MZ,Ahmad F,Kondapi AK,Qureshi IA,Ghazi IA.Antioxidant and antiproliferative activities of Abrus precatorius leaf extracts-An in vitro study.BMC Complement Altern Med 2013;13(53):1-12.
[13]Sofi MS,Sateesh MK,Bashir M,Harish G,Lakshmeesha TR,Vedashree S,et al,Cytotoxic and pro-apoptotic effects of Abrus precatorius L.on human metastatic breast cancer cell line,MDA-MB-231.Cytotechnology2013;65(3):407-417.
[14]Geran R,Greenberg N,MacDonald M,Schumacher A,Abbott256B.National Cancer Institute protocols for screening of anticancer compounds.Cancer Chemother Rep 1972;3:1-103.
[15]Igarashi M,Miyazawa T.The growth inhibitory effect of conjugated linoleic acid on a human hepatoma cell line,HepG2,is induced by a change in fatty acid metabolism,but not the facilitation of lipid peroxidation in the cells.Biochim Biophys Acta 2001;1530(2-3):162-171.
[16]Sharma A,Kaur R,Katnoria JK,Kaur R,Nagpal AK.Family Fabaceae:A boon for cancer therapy.In:Malik S(ed.)Biotechnology and production of anti-cancer compounds.Springer;2017,p.157-175.
[17]Rungruangmaitree R,Jiraungkoorskul W.Pea,Pisum sativum,and its anticancer activity.Phcog Rev 2017;11(21):39-42.
[18]Lee JH,Kim KS,Lee YC,Cho CW,Kim KM,Chung YS.Anticancer composition containing herbal extract.US 2011/0293754 A1(Patent)2017.
[19]Ahmad F,Anwar F,Hira S.Review on medicinal importance of fabaceae family.Ph OL 2016;3:151-156.
[20]Kim SJ,Min HY,Chung HJ,Park EJ,Hong JY,Kang YJ,et al.Inhibition of cell proliferation through cell cycle arrest and apoptosis by thio-ClIB-MECA,a novel A3 adenosine receptor agonist,in human lung cancer cells.Cancer Lett 2008;264(2):309-315.
[21]Li A,Blow JJ.The origin of CDK regulation.Nat Cell Biol 2001;3(8):E182-184.
[22]Czabotar PE,Lessene G,Strasser A,Adams JM.Control of apoptosis by the BCL-2 protein family:Implications for physiology and therapy.Nat Rev Mol Cell Biol 2014;15(1):49-63.
[23]Bai L,Wang S.Targeting apoptosis pathways for new cancer therapeutics.Annu Rev Med 2014;65:139-155.
[24]Chien SY,Wu YC,Chung JG,Yang JS,Lu HF,Tsao MF,et al.Quercetininduced apoptosis acts through mitochondrial-and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells.Hum Exp Toxicol2009;28(8):493-503.
[25]Pepper C,Hoy T,Bentley P.Elevated Bcl-2/Bax are a consistent feature of apoptosis resistance in B-cell chronic lymphocytic leukaemia and are correlated with in vivo chemoresistance.Leuk Lymphoma 1998;28(3-4):355-361.
[26]Beroukhim R,Mermei CH,Porter D,Wei G,Raychaudhuri S,Donovan J,et al.The landscape of somatic copy-number alteration across human cancers.Nature 2010;463(7283):899-905.
[27]Castle VP,Heidelberger KP,Bromberg J,Ou X,Dole M,Nu?ez G.Expression of the apoptosis-suppressing protein bcl-2,in neuroblastoma is associated with unfavorable histology and N-myc amplification.Am JPathol 1993;143(6):1543-1550.
[28]Olopade OI,Adeyanju MO,Safa AR,Hagos F,Mick R,Thompson CB,et al.Overexpression of BCL-x protein in primary breast cancer is associated with high tumor grade and nodal metastases.Cancer J Sci Am1997;3(4):230-237.
[29]Tait SW,Green DR.Mitochondria and cell death:Outer membrane permeabilization and beyond.Nature Rev Mol Cell Biol 2010;11(9):621-632.
[30]McIlwain DR,Berger T,Mak TW.Caspase functions in cell death and disease.Cold Spring Harb Perspect Biol 2013;5(4):a008656.Doi:10.1101/cshperspect.a008656.