, Volume 70, Issue 3, pp 913–919 | Cite as

Synergistic tumor suppression by a Perilla frutescens-derived methoxyflavanone and anti-cancer tyrosine kinase inhibitors in A549 human lung adenocarcinoma

  • Amer Ali Abd El-Hafeez
  • Takashi Fujimura
  • Rikiya Kamei
  • Noriko Hirakawa
  • Kenji Baba
  • Kazuhisa Ono
  • Seiji KawamotoEmail author
Original Article


Anti-cancer tyrosine kinase inhibitors (TKIs) are effective in many types of cancers including non-small cell lung cancer, while appearance of TKI-resistant tumors suggests a need for the development of their potentiation strategies. We have previously shown that a methoxyflavanone derivative from the Asian medicinal herb Perilla frutescens (Perilla-derived methoxyflavanone; PDMF) shows a prominent anti-tumor activity against A549 human lung adenocarcinoma. Here we show that PDMF and anti-cancer TKIs (nilotinib, bosutinib, dasatinib, and ponatinib) synergistically suppress proliferation of A549 cells. Flow cytometric analysis indicated that co-stimulation with nilotinib (4 μM) and PDMF induced G2/M cell cycle arrest in low PDMF doses (10–50 μM), whereas this combination triggered de novo G1 arrest in higher PDMF dosages (50–125 μM). We also found that co-administration with nilotinib and PDMF significantly suppressed in vivo tumorigenicity of A549 cells in athymic nude mice.


A549 cells Lung cancer Methoxyflavanone Perilla frutescens Tyrosine kinase inhibitors 



This work was financially supported by the Mishima Food Co., Ltd (to S. Kawamoto). N. Hirakawa and K. Baba are employees of the Mishima Food Co., Ltd. A. A. Abd El-Hafeez was supported by the Ministry of Education, Culture, Sports, Science, and Technology, MEXT, Japan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Supplementary material 1 (PPTX 1599 kb)
10616_2017_124_MOESM2_ESM.docx (17 kb)
Supplementary material 2 (DOCX 17 kb)


  1. Cao H, Yu S, Chen D, Jing C, Wang Z, Ma R, Liu S, Ni J, Feng J, Wu J (2016) Liver X receptor agonist T0901317 reverses resistance of A549 human lung cancer cells to EGFR-TKI treatment. FEBS Open Bio 7:35–43CrossRefPubMedPubMedCentralGoogle Scholar
  2. Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Reg 22:27–55CrossRefGoogle Scholar
  3. Garg AK, Buchholz TA, Aggarwal BB (2005) Chemosensitization and radiosensitization of tumors by plant polyphenols. Antioxid Redox Signal 7:1630–1647CrossRefPubMedGoogle Scholar
  4. Ghorbani A, Nazari M, Jeddi-Tehrani M, Zand H (2012) The citrus flavonoid hesperidin induces p53 and inhibits NF-κB activation in order to trigger apoptosis in NALM-6 cells: involvement of PPARγ-dependent mechanism. Eur J Nutr 51:39–46CrossRefPubMedGoogle Scholar
  5. Graf BA, Milbury PE, Blumberg JB (2005) Flavonols, flavones, flavanones, and human health: epidemiological evidence. J Med Food 8:281–290CrossRefPubMedGoogle Scholar
  6. Gridelli C, De Marinis F, Cappuzzo F, Di Maio M, Hirsch FR, Mok T, Morgillo F, Rosell R, Spigel DR, Yang JC, Ciardiello F (2014) Treatment of advanced non–small-cell lung cancer with epidermal growth factor receptor (EGFR) mutation or ALK gene rearrangement: results of an international expert panel meeting of the Italian Association of Thoracic Oncology. Clin Lung Cancer 15:173–181CrossRefPubMedGoogle Scholar
  7. Ito K, Semba T, Uenaka T, Wakabayashi T, Asada M, Funahashi Y (2014) Enhanced anti-angiogenic effect of E7820 in combination with erlotinib in epidermal growth factor receptor-tyrosine kinase inhibitor-resistant non-small-cell lung cancer xenograft models. Cancer Sci 105:1023–1031CrossRefPubMedPubMedCentralGoogle Scholar
  8. Jin CY, Park C, Lee JH, Chung KT, Kwon TK, Kim GY, Choi BT, Choi YH (2009) Naringenin-induced apoptosis is attenuated by Bcl-2 but restored by the small molecule Bcl-2 inhibitor, HA 14-1, in human leukemia U937 cells. Toxicol In Vitro 23:259–265CrossRefPubMedGoogle Scholar
  9. Kamei R, Fujimura T, Matsuda M, Kakihara K, Hirakawa N, Baba K, Ono K, Arakawa K, Kawamoto S (2017) A flavanone derivative from the Asian medicinal herb (Perilla frutescens) potently suppresses IgE-mediated immediate hypersensitivity reactions. Biochem Biophys Res Commun 483:674–679CrossRefPubMedGoogle Scholar
  10. Kim YA, Kim H, Seo Y (2013) Antiproliferative effect of flavonoids from the halophyte Vitex rotundifolia on human cancer cells. Nat Prod Commun 8:1405–1408PubMedGoogle Scholar
  11. Krause DS, Van Etten RA (2005) Tyrosine kinases as targets for cancer therapy. N Engl J Med 353:172–187CrossRefPubMedGoogle Scholar
  12. Kuno T, Tsukamoto T, Hara A, Tanaka T (2012) Cancer chemoprevention through the induction of apoptosis by natural compounds. J Biophys Chem 3:19431CrossRefGoogle Scholar
  13. Liu C, Wang Y, Xie S, Zhou Y, Ren X, Li X, Cai Y (2011) Liquiritigenin induces mitochondria-mediated apoptosis via cytochrome c release and caspases activation in HeLa Cells. Phytother Res 25:277–283PubMedGoogle Scholar
  14. Meoni G, Cecere FL, Lucherini E, Di Costanzo F (2013) Medical treatment of advanced non-small cell lung cancer in elderly patients: a review of the role of chemotherapy and targeted agents. J Geriatr Oncol 4:282–290CrossRefPubMedGoogle Scholar
  15. Milano V, Piao Y, LaFortune T, de Groot J (2009) Dasatinib-induced autophagy is enhanced in combination with temozolomide in glioma. Mol Cancer Ther 8:394–406CrossRefPubMedGoogle Scholar
  16. Molina JR, Yang P, Cassivi SD, Schild SE, Adjei AA (2008) Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc 5:584–594CrossRefGoogle Scholar
  17. Nautiyal J, Kanwar SS, Yu Y, Majumdar AP (2011) Combination of dasatinib and curcumin eliminates chemo-resistant colon cancer cells. J Mol Signal 6:7CrossRefPubMedPubMedCentralGoogle Scholar
  18. Park HJ, Kim MJ, Ha E, Chung JH (2008a) Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNU-C4. Phytomedicine 15:147–151CrossRefPubMedGoogle Scholar
  19. Park JH, Jin CY, Lee BK, Kim GY, Choi YH, Jeong YK (2008b) Naringenin induces apoptosis through downregulation of Akt and caspase-3 activation in human leukemia THP-1 cells. Food Chem Toxicol 46:3684–3690CrossRefPubMedGoogle Scholar
  20. Rosell R, Moran T, Queralt C, Porta R, Cardenal F, Camps C, Majem M, Lopez-Vivanco G, Isla D, Provencio M, Insa A, Massuti B, Gonzalez-Larriba JL, Paz-Ares L, Bover I, Garcia-Campelo R, Moreno MA, Catot S, Rolfo C, Reguart N, Palmero R, Sánchez JM, Bastus R, Mayo C, Bertran-Alamillo J, Molina MA, Sanchez JJ, Taron M, Spanish Lung Cancer Group (2009) Screening for epidermal growth factor receptor mutations in lung cancer. N Engl J Med 361:958–967CrossRefPubMedGoogle Scholar
  21. Sak K (2012) Chemotherapy and dietary phytochemical agents. Chemother Res Pract 2012:282570PubMedPubMedCentralGoogle Scholar
  22. Siegel R, Naishadham D, Jemal A (2013) Cancer statistics, 2013. CA Cancer J Clin 63:11–30CrossRefPubMedGoogle Scholar
  23. Surh YJ (2003) Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 3:768–780CrossRefPubMedGoogle Scholar
  24. Szliszka E, Czuba ZP, Jernas K, Król W (2008) Dietary flavonoids sensitize HeLa cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Int J Mol Sci 9:56–64CrossRefPubMedPubMedCentralGoogle Scholar
  25. Tomayko MM, Reynolds CP (1989) Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol 24:148–154CrossRefPubMedGoogle Scholar
  26. Zhang P, Gao WY, Turner S, Ducatman BS (2003) Gleevec (STI-571) inhibits lung cancer cell growth (A549) and potentiates the cisplatin effect in vitro. Mol Cancer 2:1CrossRefPubMedPubMedCentralGoogle Scholar
  27. Zhang SP, Zhou YJ, Liu Y, Cai YQ (2009) Effect of liquiritigenin, a flavanone existed from Radix glycyrrhizae on pro-apoptotic in SMMC-7721 cells. Food Chem Toxicol 47:693–701CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Hiroshima Research Center for Healthy Aging (HiHA), Department of Molecular Biotechnology, Graduate School of Advanced Sciences of MatterHiroshima UniversityHigashi-HiroshimaJapan
  2. 2.Mishima Food Co. Ltd.HiroshimaJapan
  3. 3.Department of Food Sciences and Biotechnology, Faculty of Life SciencesHiroshima Institute of TechnologyHiroshimaJapan

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