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2,3,5-Trimethoxy-4-cresol, an anti-metastatic constituent from the solid-state cultured mycelium of Antrodia cinnamomea and its mechanism


Antrodia cinnamomea is a valuable and unique edible fungus originating from the forests of Taiwan. In this study, an anti-metastatic compound, 2,3,5-trimethoxy-4-cresol (TMC), was isolated from the solid-state cultured mycelium of A. cinnamomea. According to the results obtained from cell wound healing, cell migration and invasion assays, TMC effectively suppressed movement, migration and invasion of lung cancer cells at the dosage of 5–40 μM, which was non-toxic to A549 cells. In addition, TMC reduced protein expression of Akt, MMP-2 and MMP-9 and enhanced E-cadherin and TIMP-1 protein expression, which are known to regulate cell adhesion, migration and invasion. Taken together, TMC effectively suppresses movement, migration and invasion of lung cancer cells, and achieves an anti-cancer metastasis effect.

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Dimethyl sulfoxide


Dulbecco’s phosphate-buffered saline


Extracellular matrix


Epithelial–mesenchymal transition


Focal adhesion kinase


Fetal bovine serum


Gallic acid


Matrix metalloproteinase


3-(4,5-Dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide


Non-small-cell lung cancer


Sodium dodecyl sulfate




Tissue inhibitor of metalloproteinase-2


Tumor necrosis factor alpha


  1. Simon GR (2014) nab-Paclitaxel for the treatment of advanced squamous none small-cell lung cancer: a comprehensive update. Clin Lung Cancer 15:391–397

    CAS  Article  PubMed  Google Scholar 

  2. Riihimäki M, Thomsen H, Hemminki A, Sundquist K, Hemminki K (2013) Comparison of survival of patients with metastases from known versus unknown primaries: survival in metastatic cancer. BMC Cancer 13:36

    PubMed Central  Article  PubMed  Google Scholar 

  3. Nichols L, Saunders R, Knollmann FD (2012) Causes of death of patients with lung cancer. Arch Pathol Lab Med 136:1552–1557

    Article  PubMed  Google Scholar 

  4. Wang HC, Chu FH, Chian SC, Liao JW, Hsieh HW, Li WH, Lin CC, Shaw JF, Kuo YH, Wang SY (2012) Establishment of the metabolite profile for an Antrodia cinnamomea health food product and investigation of its chemoprevention activity. J Agric Food Chem 61:8556–8564

    Article  Google Scholar 

  5. Hsieh YH, Chu FH, Wang YS, Chien SC, Chang ST, Shaw JF, Chen CY, Hsiao WW, Kuo YH, Wang SY (2010) Antrocamphin A, an anti-inflammatory principal from the fruiting body of Taiwanofungus camphoratus, and its mechanisms. J Agric Food Chem 58:3153–3158

    CAS  Article  PubMed  Google Scholar 

  6. Gokila Vani M, Senthil Kumar KJ, Liao JW, Chien SC, Mau JL, Chiang SS, Lin CC, Kuo YH, Wang SY (2013) Antcin C from Antrodia cinnamomea protects liver cells against free radical-induced oxidative stress and apoptosis in vitro and in vivo through Nrf2-dependent mechanism. Evid Based Complement Alternat Med 2013:296082

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  7. Lin TY, Chen CY, Chien SC, Hsiao WW, Chu FH, Li WH, Lin CC, Shaw JF, Wang SY (2011) Metabolite profiles for Antrodia cinnamomea fruiting bodies harvested at different culture ages and substrates from different wood. J Agric Food Chem 59:7626–7635

    CAS  Article  PubMed  Google Scholar 

  8. Senthil Kumar KJ, Chu FH, Hsieh HW, Liao JW, Li WH, Lin CC, Shaw JF, Wang SY (2011) Antroquinonol from ethanolic extract of mycelium of Antrodia cinnamomea protects hepatic cells from ethanol-induced oxidative stress through Nrf-2 activation. J Ethnopharmacol 136:168–177

    Article  Google Scholar 

  9. Lin TY, Chien SC, Kuo YH, Wang SY (2012) Distinguishing between R- and S-antcin C and their cytotoxicity. Nat Prod Commun 7:835–836

    CAS  PubMed  Google Scholar 

  10. Lu MC, El-Shazly M, Wu TY, Du YC, Chang TT, Chen CF, Hsu YM, Lai KH, Chiu CP, Chang FR, Wu YC (2013) Recent research and development of Antrodia cinnamomea. Pharmacol Ther 139:124–156

    CAS  Article  PubMed  Google Scholar 

  11. Chen YC, Liu YL, Li FY, Chang CI, Wang SY, Lee KY, Li SL, Chen YP, Jinn TR, Tzen JT (2011) Antcin A, a steroid-like compound from Antrodia camphorata, exerts anti-inflammatory effect via mimicking glucocorticoids. Acta Pharmacol Sin 32:904–911

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  12. Geethangili M, Tzeng YM (2011) Review of pharmacology effects of Antrodia camphorata and its bioactivities compounds. Evid Based Complement Alternat Med 2011:212641

    PubMed Central  Article  PubMed  Google Scholar 

  13. Ao ZH, Xu ZH, Lu ZM, Xu HY, Zhang XM, Dou WF (2009) Niuchangchih (Antrodia camphorata) and its potential in treating liver diseases. J Ethnophrarmcol 121:194–212

    Article  Google Scholar 

  14. Liu FC, Lai MT, Chen YY, Lin WH, Chang SJ, Sheu MJ, Wu CH (2013) Elucidating the inhibitory mechanisms of the ethanolic extract of the fruiting body of the mushroom Antrodia cinnamomea on the proliferation and migration of murine leukemia WEHI-3 cells and their tumorigenicity in a BALB/c allograft tumor model. Phytomedicine 20:874–882

    CAS  Article  PubMed  Google Scholar 

  15. Chen YY, Chou PY, Chien YC, Wu CH, Wu TS, Sheu MJ (2012) Ethanol extracts of fruiting bodies of Antrodia cinnamomea exhibit anti-migration action in human adenocarcinoma CL1-0 cells through the MAPK and PI3 K/AKT signaling pathways. Phytomedicine 19:768–778

    CAS  Article  PubMed  Google Scholar 

  16. Yang HL, Kuo YH, Tsai CT, Huang YT, Chen SC, Chang HW, Lin E, Lin WH, Hseu YC (2011) Anti-metastatic activities of Antrodia camphorata against human breast cancer cells mediated through suppression of the MAPK signaling pathway. Food Chem Toxicol 49:290–298

    CAS  Article  PubMed  Google Scholar 

  17. Senthil Kumar KJ, Vani MG, Chueh PJ, Mau JL, Wang SY (2015) Antrodin C inhibits epithelial-to-mesenchymal transition and metastasis of breast cancer cells via suppression of Smad2/3 and β-Catenin signaling pathways. PLoS One 10:e0117111

    Article  Google Scholar 

  18. Fa KN, Yang CM, Chen PC, Lee YY, Chyau CC, Hu ML (2015) Anti-metastatic effect of antrodan, the Antrodia cinnamomea mycelia glycoprotein in lung carcinoma cells. Int J Biol Macromol 74:476–482

    CAS  Article  PubMed  Google Scholar 

  19. Shishodia S, Chaturvedi MM, Aggarwal BB (2007) Role of curcumin in cancer therapy. Curr Probl Cancer 31:243–305

    Article  PubMed  Google Scholar 

  20. Lee TH, Chen CC, Chen JJ, Liao HF, Chang HS, Sung PJ, Tseng MH, Wang SY, Ko HH, Kuo YH (2014) New cytotoxic components from Antrodia camphorate. Molecules 19:21378–21385

    Article  PubMed  Google Scholar 

  21. Dauer DJ, Ferraro B, Song L, Yu B, Mora L, Buettner R, Enkemann S, Jove R, Haura EB (2005) Stat3 regulates genes common to both wound healing and cancer. Oncogene 24:3397–3408

    CAS  Article  PubMed  Google Scholar 

  22. Albelda S (1993) Role of integrins and other cell adhesion molecules in tumor progression and metastasis. Lab Invest J Tec Meth Pathology 68:4–17

    CAS  Google Scholar 

  23. Chen HW, Lee JY, Huang JY, Wang CC, Chen WJ, Su SF, Huang CW, Ho CC, Chen JJW, Tsai MF, Yu SL, Yang PC (2008) Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppression HLJ1. Cancer Res 68:7428–7438

    CAS  Article  PubMed  Google Scholar 

  24. Chen QY, Zheng Y, Jiao DM, Chen FY, Hu HZ, Wu YQ, Song J, Yan J, Wu LJ, Lv GY (2014) Curcumin inhibits lung cancer cell migration and invasion through Rac1-dependent signaling pathway. J Nutr Biochem 25:177–185

    Article  PubMed  Google Scholar 

  25. Kessenbrock K, Plaks V, Werb Z (2010) Matrix metalloproteinases: regulators of the tumor microenviroment. Cell 141:52–67

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  26. Gialeli C, Theocharis AD, Karamanos NK (2010) Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J 278:16–27

    Article  PubMed  Google Scholar 

  27. Roy DM, Walsh LA (2014) Candidate prognostic markers in breast cancer: focus on extracellular proteases and their inhibitors. Breast Cancer 6:81–91

    PubMed Central  CAS  PubMed  Google Scholar 

  28. Thiery JP (2002) Epithelial–mesenchymal transitions in tumour progression. Nat Rev Cancer 2:442–454

    CAS  Article  PubMed  Google Scholar 

  29. Grille SJ, Bellacosa A, Upson J, Klein-Szanto AJ, van Roy F, Lee-Kwon W, Donowitz M, Tsichlis PN, Larue L (2003) The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines. Cancer Res 63:2172–2178

    CAS  PubMed  Google Scholar 

  30. Xiao D, He J (2010) Epithelial mesenchymal transition and lung cancer. J Thoracic Disease 2:154–159

    CAS  Google Scholar 

  31. Tania M, Khan MA, Fu J (2014) Epithelial to mesenchymal transition inducing transcription factors and metastatic cancer. Tumour Biol 35:7335–7342

    CAS  Article  PubMed  Google Scholar 

  32. Zetter BR (1993) Adhesion molecules in tumor metastasis. Semin Cancer Biol 4:219–229

    CAS  PubMed  Google Scholar 

  33. Lin CT, Senthil Kumar KJ, Tseng YH, Wang ZJ, Pan MY, Xiao JH, Chien SC, Wang SY (2009) Anti-inflammatory activity of Flavokawain B from Alpinia pricei Hayata. J Agric Food Chem 57:6060–6065

    CAS  Article  PubMed  Google Scholar 

  34. Wu YY, Peck K, Chang YL, Pan SH, Cheng YF, Lin JC, Yang RB, Hong TM, Yang PC (2011) SCUBE3 is an endogenous TGF-β receptor ligand and regulates the epithelial-mesenchymal transition in lung cancer. Oncogene 30:3682–3693

    CAS  Article  PubMed  Google Scholar 

  35. Meng XN, Jin Y, Yu Y, Bai J, Liu GY, Zhu J, Zhao YZ, Wang Z, Chen F, Lee KY, Fu SB (2009) Characterisation of fibronectin-mediated FAK signalling pathways in lung cancer cell migration and invasion. Br J Cancer 101:327–334

    PubMed Central  CAS  Article  PubMed  Google Scholar 

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Parts of this study were supported by the Ministry of Science and Technology, Taiwan, Republic of China (NSC-103-2911-I-005-301, NSC-102-2911-I-005-301) and the Ministry of Education, Taiwan, R.O.C. under the ATU plan.

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No competing financial interests exist.

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Correspondence to Jong-Tar Kuo or Sheng-Yang Wang.

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Lin, CC., Chen, CC., Kuo, YH. et al. 2,3,5-Trimethoxy-4-cresol, an anti-metastatic constituent from the solid-state cultured mycelium of Antrodia cinnamomea and its mechanism. J Nat Med 69, 513–521 (2015).

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  • Antrodia cinnamomea
  • 2,3,5-Trimethoxy-4-cresol
  • Anti-metastasis
  • EMT