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Journal of Integrative Medicine ›› 2014, Vol. 12 ›› Issue (2): 102-114.doi: 10.1016/S2095-4964(14)60015-7

• Research Article • Previous Articles     Next Articles

Oleanolic acid isolated from ethanolic extract of Phytolacca decandra induces apoptosis in A375 skin melanoma cells: Drug-DNA interaction and signaling cascade

Samrat Ghosh, Kausik Bishayee, Anisur Rahman Khuda-Bukhsh   

  1. Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani-741235, West Bengal, India
  • Received:2013-11-26 Accepted:2014-01-03 Online:2014-03-10 Published:2018-10-22

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Objective

Oleanolic acid (OA) has been reported to have anticancer effects, but the extent of its cytotoxicity, its ability to interact with nuclear DNA, its action against skin melanoma, as well as the molecular mechanism of its action against cell proliferation and in support of cell death are still unexplored. This led us to examine the efficacy of OA, a bioactive compound isolated from Phytolacca decandra, on these issues in the present investigation. 

Methods

Studies related to analyses of cell viability, drug-DNA interaction, cell proliferation, cell cycle and epidermal growth factor receptor (EGFR) activity were performed. To investigate whether cells undergo apoptosis, studies like fluorescence microscopy, poly (ADP-ribose) polymerase (PARP) degradation, annexin V-fluorescein isothiocyanate/propidium iodide assay, alteration in mitochondrial membrane potential and activity of some relevant signaling proteins were performed. 

Results

OA displayed a minimal and negligible cytotoxic effect on normal HaCaT cells (skin keratinocytes) and peripheral blood mononuclear cells but by contrast it reduced A375 cell viability significantly. OA interacted with nuclear DNA quickly after exposure. It acted as an anti-proliferative agent. It suppressed EGFR activity. OA administration led the cells to mitochondria-dependent caspase 3-mediated apoptosis. 

Conclusion

OA interacts with cellular DNA, inhibits proliferation possibly through modulating EGFR activity and induces mitochondria-dependent caspase 3-mediated apoptosis in A375 cells which would qualify it as a potent anticancer agent.

Key words: Oleanolic acid, Skin melanoma, Drug-DNA interaction, Proliferation, Epidermal growth factor receptor, Apoptosis

Figure 1

Mass spectra and 1H NMR (A) Mass spectra of oleanolic acid isolated from ethanolic extract of Phytolacca decandra. Mass spectroscopy data revealed that the molecular weight of isolated compound from ethanolic extract of P. decandra was m/z: 479.73 [M+23]+, which implied that the actual molecular weight of the compound was approximately (479.73-23.00) or 456.73. (B) 1H NMR of oleanolic acid isolated from ethanolic extract of P. decandra. 1H NHR: 1H nuclear magnetic resonance."

Figure 2

FTIR spectra, molecular structure of OA and cell viability assay (A) FTIR spectra of OA isolated from ethanolic extract of Phytolacca decandra. (B) Molecular structure of OA. (C) Cell viability assay. Cell viabilities were assessed by using MTT assay. A375, HaCaT and PBMCs were exposed to different concentrations of OA (0-48.17 μmol/L) for 24 h. The values of three independent experiments were represented as mean ± standard error of mean. Statistical significance was considered as P<0.05. *P<0.05, vs relevant control. FTIR: Fourier transform infrared; OA: oleanolic acid; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PBMCs: peripheral blood mononuclear cells."

Figure 3

Drug-DNA interaction study, monolayer growth assay and cell cycle assayDNA binding capacity of OA was determined by CD spectroscopy. (A) Interaction with CT-DNA. The CT-DNA (100 μmol/L) was exposed to 40.7 μmol/L OA. Uppermost curve indicated in black represents vehicle-treated CT-DNA and the ash coloured curve represents OA-treated CT-DNA. (B) Interaction with cellular DNA. A375 cells were treated with 40.7 μmol/L of OA for 3 h. Thereafter, cellular DNA was isolated, and purified and interaction was checked. (C) Thermal denaturation study of CT-DNA: thermal denaturation of CT-DNA (30 μg/mL) in absence (triangles) and presence (circles) of OA (40.7 μmol/L). (D) Monolayer growth assay. Treatment with 40.70 μmol/L of OA reduced the proliferation of the A375 cells in a time-dependent manner at late hours of treatment. The bars represented the fractional value of cell count, considering the cell count of control as unity. Statistical significance was considered as P<0.05. *P<0.05, vs relevant control. (E) Cell cycle analysis. Propidium iodide-based FACS (FL2 filter) assay was performed to analyze the cell cycle progression. The percentages of cells of various phases have been mentioned. OA: oleanolic acid; CT-DNA: calf thymus-DNA."

Figure 4

EGFR activity assay and activity study on its downstream signaling proteins EGFR activity has been assessed by immunoblot analysis following treatment with OA (40.70 μmol/L) for 3 h and 6 h in presence or absence of EGF (10 nmol/L). Cells were cultured in serum-free medium for 12 h before being treated with EGF only or with EGF and OA. (A) Immunoblotting pictures of EGFR and pEGFR. (B) The corresponding densitometric analysis of intensities of immunoblots. Statistical significance was considered as P<0.05. *P<0.05, vs control for EGFR 3 h; △P<0.05, vs control for EGFR 6 h; ▲P<0.05, vs control for pEGFR 3 h; □P<0.05, vs control for pEGFR 6 h, respectively. (C) Immunoblotting pictures of pAkt, Erk1/2, and pSTAT5 after exposing the A375 cells to OA (D1 = 20 μmol/L, D2 = 30 μmol/L, D3 = 40.7 μmol/L) for 24 h. (D) The corresponding densitometric analysis of intensities of immunoblots has been plotted. Statistical significance was considered as P<0.05. *P<0.05, vs control. EGFR: epidermal growth factor receptor; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 5

Apoptosis analysis Cells were treated with OA of specified doses (D1 = 20 μmol/L, D2 = 30 μmol/L, D3 = 40.7 μmol/L) for 24 h. (A) Nucleosomal fragmentation analysis. OA-treated cells showed DAPI-positive whereas control cells were DAPI-negative. Magnification was 200×. (B) PARP degradation study by immunoblot assay. The densitometric plot of corresponding band intensity shows the increase in intensity in a dose-dependent manner. Statistical significance was considered as P<0.05. *P<0.05, vs control. (C) Annexin V-FITC/PI dual stain assay. Assessment of externalization of phosphatidyl serine by annexin V/PI assay was done by FACS analysis. LL–: lower left (Annexin V–/PI–); UL–: upper left (Annexin V–/PI+); LR–: lower right (Annexin V+/PI–); UR–: upper right (Annexin V+/PI+). The corresponding percentages of cells present on those quadrants were mentioned in upper right corners. DAPI: 4′-6-diamidino-2-phenylindole; FITC: fluorescein isothiocyanate; PI: propidium iodide; PARP: poly (ADP-ribose) polymerase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 6

MMP alteration detection, immunoblotting of some signaling proteins and caspase 3 activity assay (A) Detection of MMP alteration. Change in mitochondrial membrane potential was evaluated by FACS analysis, after treating the cells with 40.70 μmol/L of OA for 12, 18, and 24 h. The shifting of fluorescence peak in the overlay histogram, obtained from flow cytometric data (using FL1 filter), was towards the y-axis and the shifting was most at the 24th hour of OA treatment. ‘M’ represents an arbitrary area along the rhodamine 123 fluorescence axis. The cell populations (within the area of ‘M’) are calculated in percentages and mentioned in the figure. (B) Immunoblotting. Cells were treated for 24 h with specified doses (D1 = 20 μmol/L, D2 = 30 μmol/L, D3 = 40.7 μmol/L). GAPDH was served as an equal loading control. The densitometric plot of relative band intensities revealed that the regulation was dose-dependent. (C) Caspase 3 activity assay. Caspase 3 activity was detected by immunoblot assay. Cells were treated with 40.70 μmol/L of OA only for 24 h or treated with 40.70 μmol/L of OA for 24 h after pre-treated with z-DEVD-fmk (caspase 3 inhibitor, 10 μmol/L) for 2 h. Relative band intensities of cleaved caspase 3 and cleaved PARP fragment revealed up-regulated expression of both upon treatment with OA only whereas substantial inhibition of caspase 3 and cleaved PARP levels was observed following pre-treatment with caspase 3 inhibitor with respect to cells treated with OA only. Statistical significance was considered as P<0.05. *P<0.05, vs control. MMP: mitochondrial membrane potential; OA: oleanolic acid; PARP: poly (ADP-ribose) polymerase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

[1] Palchaudhuri R, Hergenrother PJ . DNA as a target for anticancer compounds: methods to determine the mode of binding and the mechanism of action[J]. Curr Opin Biotechnol, 2007,18(6):497-503
doi: 10.1016/j.copbio.2007.09.006
[2] Plesca D, Mazumder S, Almasan A . DNA damage response and apoptosis[J]. Methods Enzymol, 2008,446:107-122
doi: 10.1016/S0076-6879(08)01606-6
[3] Raff MC . Social controls on cell survival and cell death[J]. Nature, 1992,356(6368):397-400
doi: 10.1038/356397a0 pmid: 1557121
[4] Yarden Y, Sliwkowski MX . Untangling the ErbB signalling network[J]. Nat Rev Mol Cell Biol, 2001,2(2):127-137
doi: 10.1038/35052073 pmid: 11252954
[5] Alroy I, Yarden Y . The ErbB signaling network in embryogenesis and oncogenesis: signal diversification through combinatorial ligand-receptor interactions[J]. FEBS Lett, 1997,410(1):83-86
doi: 10.1016/S0014-5793(97)00412-2
[6] Schlessinger J . Cell signaling by receptor tyrosine kinases[J]. Cell, 2000,103(2):211-225
doi: 10.1016/j.cell.2010.06.011 pmid: 2060299620602996
[7] Boone B, Jacobs K, Ferdinande L, Taildeman J, Lambert J, Peeters M, Bracke M, Pauwels P, Brochez L . EGFR in melanoma: clinical significance and potential therapeutic target[J]. J Cutan Pathol, 2011,38(6):492-502
doi: 10.1111/j.1600-0560.2011.01673.x pmid: 21352258
[8] Dzubak P, Hajduch M, Vydra D, Hustova A, Kvasnica M, Biedermann D, Markova L, Urban M, Sarek J . Pharmacological activities of natural triterpenoids and therapeutic implications[J]. Nat Prod Rep, 2006,23(3):394-411
doi: 10.1002/chin.200637244 pmid: 16741586
[9] Liu J . Pharmacology of oleanolic acid and ursolic acid[J]. J Ethnopharmacol, 1995,49(2):57-68
doi: 10.1016/0378-8741(95)90032-2 pmid: 8847885
[10] Jeong TS, Hwang EI, Lee HB, Lee ES, Kim YK, Min BS, Bae KH, Bok SH, Kim SU . Chitin synthase II inhibitory activity of ursolic acid, isolated from Crataegus pinnatifida[J]. Planta Med, 1999,65(3):261-263
doi: 10.1055/s-2006-960474
[11] Tang HQ, Hu J, Yang L, Tan RX . Terpenoids and flavonoids from Artemisia species[J]. Planta Med, 2000,66(4):391-393
doi: 10.1055/s-2000-8538 pmid: 10865468
[12] Marquina S, Maldonado N , Gardu?o-Ramírez ML, Aranda E, Villarreal ML, Navarro V, Bye R, Delgado G, Alvarez L.Bioactive oleanolic acid saponins and other constituents from the roots of Viguiera decurrens[J]. Phytochemistry, 2001,56(1):93-97
doi: 10.1016/S0031-9422(00)00283-1 pmid: 11198824
[13] Kashiwada Y, Nagao T, Hashimoto A, Ikeshiro Y, Okabe H, Cosentino LM, Lee KH . Anti-AIDS agents 38. Anti-HIV activity of 3-O-acyl ursolic acid derivatives[J]. J Nat Prod, 2000,63(12):1619-1622
doi: 10.1021/np990633v
[14] Ma C, Nakamura N, Hattori M, Kakuda H, Qiao J, Yu H . Inhibitory effects on HIV-1 protease of constituents from the wood of Xanthoceras sorbifolia[J]. J Nat Prod, 2000,63(2):238-242
doi: 10.1021/np9902441
[15] Alvarez ME, Maria AO, Saad JR . Diuretic activity of Fabiana patagonica in rats[J]. Phytother Res, 2002,16(1):71-73
doi: 10.1002/ptr.754 pmid: 11807970
[16] Assefa H, Nimrod A, Walker L, Sindelar R . Enantioselective synjournal and complement inhibitory assay of A/B-ring partial analogues of oleanolic acid[J]. Bioorg Med Chem Lett, 2001,11(13):1619-1623
doi: 10.1016/S0960-894X(01)00210-4
[17] Li J, Xu LZ, Zhu WP, Zhang TM, Li XM, Jin AP, Huang KM, Li DL, Yang QY . Effects of ursolic acid and oleanolic acid on Jurkat lymphoma cell line in vitro[J]. Zhongguo Ai Zheng Za Zhi, 1999,9(5-6):395-397
[18] Baricevic D, Sosa S, Della Loggia R, Tubaro A, Simonovska B, Krasna A, Zupancic A . Topical anti-inflammatory activity of Salvia officinalis L. leaves: the relevance of ursolic acid[J]. J Ethnopharmacol, 2001,75(2-3):125-132
doi: 10.1016/S0378-8741(00)00396-2
[19] Giner-Larza EM , Má?ez S, Recio MC, Giner RM, Prieto JM, Cerdá-Nicolás M, Ríos JL.Oleanonic acid, a 3-oxotriterpene from Pistacia, inhibits leukotriene synjournal and has anti-inflammatory activity[J]. Eur J Pharmacol, 2001,428(1):137-143
doi: 10.1016/S0014-2999(01)01290-0
[20] Ismaili H, Tortora S, Sosa S, Fkih-Tetouani S, Ilidrissi A, Della Loggia R, Tubaro A, Aquino R . Topical anti-inflammatory activity of Thymus willdenowii[J]. J Pharm Pharmacol, 2001,53(12):1645-1652
doi: 10.1211/0022357011778250 pmid: 11804395
[21] Ryu SY, Oak MH, Yoon SK, Cho DI, Yoo GS, Kim TS, Kim KM . Anti-allergic and anti-inflammatory triterpenes from the herb of Prunella vulgaris[J]. Planta Med, 2000,66(4):358-360
doi: 10.1055/s-2000-8531 pmid: 10865455
[22] Janakiram NB, Indranie C, Malisetty SV, Jagan P, Steele VE, Rao CV . Chemoprevention of colon carcinogenesis by oleanolic acid and its analog in male F344 rats and modulation of COX-2 and apoptosis in human colon HT-29 cancer cells[J]. Pharm Res, 2008,25(9):2151-2157
doi: 10.1007/s11095-008-9582-7
[23] Juan ME, Planas JM, Ruiz-Gutierrez V, Daniel H, Wenzel U . Antiproliferative and apoptosis-inducing effects of maslinic and oleanolic acids, two pentacyclic triterpenes from olives, on HT-29 colon cancer cells[J]. Br J Nutr, 2008,100(1):36-43
[24] Li J, Guo WJ, Yang QY . Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15[J]. World J Gastroenterol, 2002,8(3):493-495
doi: 10.3748/wjg.v8.i3.493 pmid: 12046077
[25] Lúcio KA, Rocha Gda G , Mon??o-Ribeiro LC, Fernandes J, Takiya CM, Gattass CR.Oleanolic acid initiates apoptosis in non-small cell lung cancer cell lines and reduces metastasis of a B16F10 melanoma model in vivo[J]. PLoS One, 2011,6(12):e28596
doi: 10.1371/journal.pone.0028596
[26] Pratheeshkumar P, Kuttan G . Oleanolic acid induces apoptosis by modulating p53, Bax, Bcl-2 and caspase-3 gene expression and regulates the activation of transcription factors and cytokine profile in B16F[J]. J Environ Pathol Toxicol Oncol, 2011,30(1):21-31
doi: 10.1615/JEnvironPatholToxicolOncol.v30.i1
[27] Shyu MH, Kao TC, Yen GC . Oleanolic acid and ursolic acid induce apoptosis in HuH7 human hepatocellular carcinoma cells through a mitochondrial-dependent pathway and downregulation of XIAP[J]. J Agric Food Chem, 2010,58(10):6110-6118
doi: 10.1021/jf100574j
[28] Tokuda H, Ohigashi H, Koshimizu K, Ito Y . Inhibitory effects of ursolic and oleanolic acid on skin tumor promotion by 12-O-tetradecanoylphorbol-13-acetate[J]. Cancer Lett, 1986,33(3):279-285
doi: 10.1016/0304-3835(86)90067-4
[29] Yan SL, Huang CY, Wu ST, Yin MC . Oleanolic acid and ursolic acid induce apoptosis in four human liver cancer cell lines[J]. Toxicol In Vitro, 2010,24(3):842-848
doi: 10.1016/j.tiv.2009.12.008 pmid: 20005942
[30] Wei J, Liu M, Liu H, Wang H, Wang F, Zhang Y, Han L, Lin X . Oleanolic acid arrests cell cycle and induces apoptosis via ROS-mediated mitochondrial depolarization and lysosomal membrane permeabilization in human pancreatic cancer cells[J]. J Appl Toxicol, 2013,33(8):756-765
doi: 10.1002/jat.v33.8
[31] Zhang P, Li H, Chen D, Ni J, Kang Y, Wang S . Oleanolic acid induces apoptosis in human leukemia cells through caspase activation and poly(ADP-ribose) polymerase cleavage[J]. Acta Biochim Biophys Sin (Shanghai), 2007,39(10):803-809
doi: 10.1111/abbs.2007.39.issue-10
[32] Ghosh S, Bishayee K, Paul A, Mukherjee A, Sikdar S, Chakraborty D, Boujedaini N, Khuda-Bukhsh AR . Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation[J]. J Integr Med, 2013,11(2):116-124
doi: 10.3736/jintegrmed2013014
[33] Ruitenberg JJ, Mulder CB, Maino VC, Landay AL, Ghanekar SA . VACUTAINER CPT and Ficoll density gradient separation perform equivalently in maintaining the quality and function of PBMC from HIV seropositive blood samples[J]. BMC Immunol, 2006,7:11
doi: 10.1186/1471-2172-7-11
[34] Bishayee K, Ghosh S, Mukherjee A, Sadhukhan R, Mondal J, Khuda-Bukhsh AR. Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interactionTHER].Bishayee K, Ghosh S, Mukherjee A, Sadhukhan R, Mondal J, Khuda-Bukhsh AR. Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interaction. Cell Prolif. 2013; 46(2) 153-163.
doi: 10.1111/cpr.2013.46.issue-2
[35] Ghosh R, Bhowmik S, Bagchi A, Das D, Ghosh S . Chemotherapeutic potential of 9-phenyl acridine: biophysical studies on its binding to DNA[J]. Eur Biophys J, 2010,39(8):1243-1249
doi: 10.1007/s00249-010-0577-z pmid: 20135310
[36] Seebacher W, Simic N, Weis R, Saf R, Kunert O . Complete assignments of 1H and 13C NMR resonances of oleanolic acid, 18α-oleanolic acid, ursolic acid and their 11-oxo derivatives[J]. Magn Reson Chem, 2003,41(8):636-638
doi: 10.1002/(ISSN)1097-458X
[37] Kelloff GJ, Johnson JR, Crowell JA, Boone CW , DeGeorge JJ, Steele VE, Mehta MU, Temeck JW, Schmidt WJ, Burke G, Greenwald P, Temple RJ.Approaches to the development and marketing approval of drugs that prevent cancer[J]. Cancer Epidemiol Biomarkers Prev, 1995,4(1):1-10
doi: 10.1007/BF00051683 pmid: 7894318
[38] Frenkel M, Cohen L . Effective communication about the use of complementary and integrative medicine in cancer care[J]. J Altern Complement Med, 2014,20(1):12-18
doi: 10.1089/acm.2012.0533
[39] Lawlor MA, Alessi DR. PKB/Akt: a key mediator of cell proliferation, survival and insulin responses? JTHER].Lawlor MA, Alessi DR. PKB/Akt: a key mediator of cell proliferation, survival and insulin responses?[J] Cell Sci. 2001; 114(pt 16):2903-2910.
[40] Gerl R, Vaux DL . Apoptosis in the development and treatment of cancer[J]. Carcinogenesis, 2005,26(2):263-270
doi: 10.1093/carcin/bgh283 pmid: 15375012
[41] Collins JA, Schandi CA, Young KK, Vesely J, Willingham MC . Major DNA fragmentation is a late event in apoptosis[J]. J Histochem Cytochem, 1997,45(7):923-934
doi: 10.1177/002215549704500702 pmid: 9212818
[42] Tsuruta F, Masuyama N, Gotoh Y . The phosphatidylinositol 3-kinase(PI3K)-Akt pathway supresses Bax translocation to mitochondria[J]. J Biol Chem, 2002,277(16):14040-14047
doi: 10.1074/jbc.M108975200
[43] Boulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M . Role of poly(ADP-ribose) polymerase(PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells[J]. J Biol Chem, 1999,274(33):22932-22940
doi: 10.1074/jbc.274.33.22932
[44] Elmore S . Apoptosis: a review of programmed cell death[J]. Toxicol Pathol, 2007,35(4):495-516
doi: 10.1080/01926230701320337 pmid: 2117903
[45] Green DR, Reed JC . Mitochondria and apoptosis[J]. Science, 1998,8(7):267-271
[46] García-Sáez AJ . The secrets of the Bcl-2 family[J]. Cell Death Differ, 2012,19(11):1733-1740
doi: 10.1038/cdd.2012.105 pmid: 3469065
[47] Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD, Wang HG, Reed JC, Nicholson DW, Alnemri ES, Green DR, Martin SJ . Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner[J]. J Cell Biol, 1999,144(2):281-292
doi: 10.1083/jcb.144.2.281
[48] Porter AG , J?nicke RU.Emerging roles of caspase-3 in apoptosis[J]. Cell Death Differ, 1999,6(2):99-104
doi: 10.1038/sj.cdd.4400476
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Journal of Integrative Medicine, ISSN: 2095-4964 CN 31-2083/R
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