, Volume 68, Issue 5, pp 2075–2081 | Cite as

Extract of Calvatia gigantea inhibits proliferation of A549 human lung cancer cells

  • Canan Eroğlu
  • Mücahit Seçme
  • Pelin Atmaca
  • Oğuzhan Kaygusuz
  • Kutret Gezer
  • Gülseren Bağcı
  • Yavuz Dodurga
Original Article


In this study, in order to investigate the anticancer mechanism of Calvatia gigantea extract, edible mushroom species, which belong to Lycoperdaceae family, changes of CCND1, CCND2, CDK4, p21, Akt, Bax, Bcl-2, p53, caspase-3 and caspase-9 were evaluated in A549 lung cancer cells. Cytotoxic effect of C. gigantea extract was evaluated by using XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxanilide). The C. gigantea extract was treated in a time and dose dependent manner within the range 25 μg/ml–2 mg/ml to determine the IC50 dose. IC50 dose for C. gigantea extract was detected as 500 μg/ml for 72 h. According to expression results, while CCND1, CCND2, CDK4, Akt and Bcl-2 expression clearly decreased, Bax, p53, caspase-3 and caspase-9 expression clearly increased in the dose group cells (A549 cells treated with 500 μg/ml dose of C. gigantea extract for 72 h). However, there was no change in p21 expression. C. gigantea extract induced cell cycle arrest and apoptosis by decreasing the CCND1, CCND2, CDK4, Akt and Bcl-2 expression and by increasing Bax, p53, caspase-3 and caspase-9 expression in A549 cells. Mushrooms are eukaryotic organisms heavily used because of their supposedly anticancer effect. Many mushroom species have been used for medical purposes, as a result of also having many effects such as antibiotic, antiviral and anticancer effects. It is thought that the C. gigantea extract may be a significant agent for treatment of lung cancer as a single agent or in combination with other drugs.


Apoptosis Calvatia gigantea Cell cycle Lung cancer cell line 


  1. Ahmad N, Adhami VM, Afaq F, Feyes DK, Mukhtar H (2001) Resveratrolcauses WAF-1/p21-mediatedG(1)-phase arrest of cell cycle and induction of apoptosis in human epidermoid carcinoma A431 cells. Clin Cancer Res 7:1466–1473Google Scholar
  2. Bloom J, Cross FR (2007) Multiple levels of cyclin specificity in cell-cycle control. Nat Rev Mol Cell Biol 8:149–160CrossRefGoogle Scholar
  3. Chen XL, Ren KH, He HW, Shao RG (2008) Involvement of PI3K/AKT/GSK3 beta pathway in tetrandrine-induced G1 arrest and apoptosis. Cancer Biol Ther 7:1073–1078CrossRefGoogle Scholar
  4. Chen A, Huang X, Xue Z, Cao D, Huang K, Chen J, Pan Y, Gao Y (2015) The role of p21 in apoptosis, proliferation, cell cycle arrest, and antioxidant activity in UVB-irradiated human HaCaT keratinocytes. Med Sci Monit Basic Res 21:86–95. doi: 10.12659/MSMBR.893608 CrossRefGoogle Scholar
  5. Chintalgattu V, Rees ML, Culver JC, Goel A, Jiffar T, Zhang J, Dunner K Jr, Pati S, Bankson JA, Pasgualini R, Arap W, Bryan NS, Taegtmeyer H, Langley RR, Yao H, Kupferman ME, Entman ML, Dickinson ME, Khakoo AY (2013) Coronary microvascular pericytes are the cellular target of sunitinib malate-induced cardiotoxicity. Sci Transl Med 5:187ra69. doi: 10.1126/scitranslmed.3005066 CrossRefGoogle Scholar
  6. De Petris L, Crino L, Scagliotti GV, Gridelli C, Galetta D, Metro G, Novello S, Maione P, Colucci G, de Marinis F (2006) Treatment of advanced non-small cell lung cancer. Ann Oncol 2:ii36–ii41Google Scholar
  7. Fuentes-Prior P, Salvesen GS (2004) The protein structures that shape caspase activity, specificity, activation and inhibition. Biochem J 384(Pt 2):201–232CrossRefGoogle Scholar
  8. Fulda S, Debatin KM (2006) Extrinsic versus intrinsic apoptosis pathway sin anticancer chemotherapy. Oncogene 25:4798–4811CrossRefGoogle Scholar
  9. Hann D, Baker F, Denniston M, Gesme D, Reding D, Flynn T, Kennedy J, Kieltyka RL (2002) The influence of social support on depressive symptoms in cancer patients: age and gender differences. J Psychosom Res 52:279–283CrossRefGoogle Scholar
  10. Hellwig CT, Passante E, Rehm M (2011) Themolecular machinery regulating apoptosis signal transduction and it simplicationin humanphysiology and pathophysiologies. Curr Mol Med 11:31–47CrossRefGoogle Scholar
  11. Hseu YC, Lee CC, Chen YC, Kumar KJS, Chen CS, Huang YC, Hsu LS, Huang HC, Yang HL (2014) The anti-tumor activity of Antrodia salmonea in human promyelocytic leukemia (HL-60) cells is mediated via the induction of G1 cell-cycle arrest and apoptosis in vitro or in vivo. J Ethnopharmacol 153:499–510CrossRefGoogle Scholar
  12. Huang GJ, Pan CH, Liu FC, Wu TS, Wu CH (2012) Anti-inflammatory effects of ethanolic extract of Antrodia salmonea in the lipopolysaccharide-stimulated RAW246.7 macrophages and the lambda-carrageenan-induced paw edema model. Food Chem Toxicol 50:1485–1493CrossRefGoogle Scholar
  13. Jayakumar T, Ramesh E, Geraldine P (2006) Antioxidant activity of the oyster mushroom, Pleurotus ostreatus on CCl4 induced liver injury in rats. Food Chem Toxicol 44:1989–1996CrossRefGoogle Scholar
  14. Jiang X, Wang X (2004) Cytochrome C-mediated apoptosis. Annu Rev Biochem 73:87–106CrossRefGoogle Scholar
  15. Jiao C, Xie YZ, Yang X, Li H, Li XM, Pan HH, Cai MH, Zhong HM, Yang BB (2013) Anticancer activity of Amauroderma rude. PLoS One 8(6):e66504CrossRefGoogle Scholar
  16. Kahlos K, Kangas L, Hiltunen R (1989) Ergosterol peroxide, an active compound from Inonotus radiatus. Planta Med 55:389–390CrossRefGoogle Scholar
  17. Khuri FR, Herbst RS, Fossella FV (2001) Emerging therapies in non-small-cell lung cancer. Ann Oncol 12:739–744CrossRefGoogle Scholar
  18. Kurashıge S, Akuzawa Y, Endo F (1997) Effects of Lentinus edodes, Grifola frondosa and Pleurotus ostreatus administration on cancer outbreak, and activities of macrophages and lymphocytes in mice treated with a carcinogen, N-butyl-N-butanolnitrosoamine. Immunopharmacol Immunotoxicol 19:175–183CrossRefGoogle Scholar
  19. Lam YW, Ng TB, Wang HX (2001) Antiproliferative and antimitogenic activities in a peptide from puffball mushroom Calvatia caelata. Biochem Biophys Res Commun 289:744–749CrossRefGoogle Scholar
  20. Li Q, Wei YQ, Wen YJ, Zhao X, Tian L, Yang L, Mao YQ, Kan B, Wu Y, Ding ZY, Deng HX, Li J, Luo Y, Li HL, He QM, Su JM, Xiao F, Zou CH, Fu CH, Xie XJ, Yi T, Tan GH, Wang L, Chen J, Liu J, Gao ZN (2004) Induction of apoptosis and tumor regression by vesicular stomatitis virus in the presence of gemcitabine in lung cancer. Int J Cancer 112:143–149CrossRefGoogle Scholar
  21. Meng J, Fan Y, Sua M, Chen C, Ren T, Wang J, Zhao Q (2014) WLIP derived from Lasiosphaera fenzlii Reich exhibits anti-tumor activity and induces cell cycle arrest through PPAR-γ associated pathways. Int Immunopharmacol 19:37–44CrossRefGoogle Scholar
  22. Miyazawa M, Ishii T, Kirinashizawa M, Yasuda K, Hino O, Hartman PS, Ishii N (2008) Cell growth of the mouse SDHC mutant cells was suppressed by apoptosis throughout mitochondrial pathway. Biosci Trends 2:22–30Google Scholar
  23. Molina JR, Adjei AA, Jett JR (2006) Advances in chemotherapy of non-small cell lung cancer. Chest 130:1211–1219CrossRefGoogle Scholar
  24. Nam KS, Jo YS, Kim YH, Hyun JW, Kim HW (2001) Cytotoxic activities of acetoxyscirpenediol and ergosterol peroxide from Paecilomyces tenuipes. Life Sci 69:229–237CrossRefGoogle Scholar
  25. Pan MH, Chen WJ, Lin Shiau SY, Ho CT, Lin JK (2002) Tangeretin induces cell cycle G1 arrest through inhibiting cyclin-dependent kinases 2 and 4 activities as well as elevating Cdk inhibitors p21 and p27 in human colorectal carcinoma cells. Carcinogenesis 23:1677–1684CrossRefGoogle Scholar
  26. Rai M, Tidke G, Wasser SP (2005) Therapeutic potential of mushrooms. Nat Prod Radiance 4:246–257Google Scholar
  27. Ren L, Hemar Y, Perera CO, Lewis G, Krissansen GW, Buchanan PK (2014) Antibacterial and antioxidant activities of aqueous extracts of eight edible mushrooms. Bioact Carbohydr Diet Fibre 3:41–51CrossRefGoogle Scholar
  28. Roland JF, Chmielewicz ZF, Weiner BA, Gross AM, Boening OP, Luck JV, Bardos TJ, Reilly HC, Sugiura K, Stock CC, Lucas EH, Byerrum RU, Scevena JA (1960) Calvacin: a new antitumor agent. Science 132:897CrossRefGoogle Scholar
  29. Sarangı I, Ghosh D, Bhutia SK, Mallick SK, Maiti TK (2006) Anti-tumor and immunomodulating effects of Pleurotus ostreatus mycelia-derived proteoglycans. Int Immunopharmacol 6:1287–1297CrossRefGoogle Scholar
  30. Siegel R, Ma J, Jemal A (2014) Cancer statistics. CA Cancer J Clin 64:9–29CrossRefGoogle Scholar
  31. Solak MH, Kalmış E, Saglam H, Kalyoncu F (2006) Antimicrobial activity of two wild mushrooms, Clitocybe alexandri (Gill.) Konr. and Rhizopogon roseolus (Corda) T.M. Fries, collected from Turkey. Phytother Res 20:1085–1087CrossRefGoogle Scholar
  32. Surh YJ (2003) Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 3:768–780CrossRefGoogle Scholar
  33. Taylor RC, Cullen SP, Martin SJ (2008) Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol 9:231–241CrossRefGoogle Scholar
  34. Tsay JG, Chung KT, Yeh CH, Chen WL, Chen CH, Lin MHC, Lu FJ, Chiou JF, Chen CH (2009) Calvatia lilacina protein-extract induces apoptosis through glutathione depletion in human colorectal carcinoma cells. J Agric Food Chem 57:1579–1588CrossRefGoogle Scholar
  35. Wang HX, Ng TB, Ooi V, Liu WK, Chang ST (1996) A polysaccharide-peptide complex from cultured mycelia of the mushroom Tricholoma mongolicum with immunoenhancing and anti-tumor activities. Biochem Cell Biol 74:95–100CrossRefGoogle Scholar
  36. Yamasaki RJ, Strasser A (2008) The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47–59CrossRefGoogle Scholar
  37. Yamasaki F, Zhang D, Bartholomeusz C, Sudo T, Hortobagyi GN, Kurisu K, Ueno NT (2007) Sensitivity of breast cancer cells to erlotinib depends on cyclin-dependent kinase 2 activity. Mol Cancer Ther 6:2168–2177CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Canan Eroğlu
    • 1
  • Mücahit Seçme
    • 1
  • Pelin Atmaca
    • 2
  • Oğuzhan Kaygusuz
    • 2
  • Kutret Gezer
    • 2
  • Gülseren Bağcı
    • 3
  • Yavuz Dodurga
    • 1
  1. 1.Department of Medical Biology, Faculty of MedicinePamukkale UniversityKınıklı/DenizliTurkey
  2. 2.Department of Biology, Faculty of SciencePamukkale UniversityDenizliTurkey
  3. 3.Department of Medical Genetics, Faculty of MedicinePamukkale UniversityDenizliTurkey

Personalised recommendations