Skip to main content
SpringerLink
Log in

Delicaflavone induces autophagic cell death in lung cancer via Akt/mTOR/p70S6K signaling pathway

  • Original Article
  • Published:
Journal of Molecular Medicine Aims and scope Submit manuscript

Abstract

Searching for potential anticancer agents from natural sources is an effective strategy for developing novel chemotherapeutic agents. In this study, data supporting the in vitro and in vivo anticancer effects of delicaflavone, a rarely occurring biflavonoid from Selaginella doederleinii, were reported. Delicaflavone exhibited favorable anticancer properties, as shown by the MTT assay and xenograft model of human non-small cell lung cancer in male BALB/c nude mice without observable adverse effect. By transmission electron microscopy with acridine orange and Cyto-ID®Autophagy detection dyes, Western blot analysis, and RT-PCR assay, we confirmed that delicaflavone induces autophagic cell death by increasing the ratio of LC3-II to LC3-I, which are autophagy-related proteins, and promoting the generation of acidic vesicular organelles and autolysosomes in the cytoplasm of human lung cancer A549 and PC-9 cells in a time- and dose-dependent manner. Delicaflavone downregulated the expression of phospho-Akt, phospho-mTOR, and phospho-p70S6K in a time- and dose-dependent manner, suggesting that it induced autophagy by inhibiting the Akt/mTOR/p70S6K pathway in A549 and PC-9 cells. Delicaflavone is a potential anticancer agent that can induce autophagic cell death in human non-small cell lung cancer via the Akt/mTOR/p70S6K signaling pathway. Delicaflavone showed anti-lung cancer effects in vitro and in vivo. Delicaflavone induced autophagic cell death via Akt/mTOR/p70S6K signaling pathway. Delicaflavone did not show observable side effects in a xenograft mouse model. Delicaflavone may represent a potential therapeutic agent for lung cancer.

Key messages

  • Delicaflavone showed anti-lung cancer effects in vitro and in vivo.

  • Delicaflavone induced autophagic cell death via Akt/mTOR/p70S6K signaling pathway.

  • Delicaflavone did not show observable side effects in a xenograft mouse model.

  • Delicaflavone may represent a potential therapeutic agent for lung cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Jemal A, Center MM, De Santis C, Ward EM (2010) Global patterns of cancer incidence and mortality rates and trends. Cancer Epidemiol Biomark Prev 19:1893–1907

    Article  Google Scholar 

  2. Mizushima N, Levine B, Cuervo AM, Klionsky DJ (2008) Autophagy fights disease through cellular self-digestion. Nature 451:1069–1075

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Rubinsztein DC (2006) The roles of intracellular protein degradation pathways in neurodegeneration. Nature 443:780–786

    Article  CAS  PubMed  Google Scholar 

  4. Mizushima N (2007) Autophagy: process and function. Genes Dev 21:2861–2873

    Article  CAS  PubMed  Google Scholar 

  5. Anding AL, Baehrecke EH (2015) Autophagy in cell life and cell death. Curr Top Dev Biol 114:67–91

    Article  PubMed  Google Scholar 

  6. Denton D, Xu TQ, Kumar S (2015) Autophagy as a pro-death pathway. Immunol Cell Biol 93:35–42

    Article  CAS  PubMed  Google Scholar 

  7. Nelson C, Baehrecke EH (2014) Eaten to death. FEBS J 281:5411–5417

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Lum JJ, Bauer DE, Kong M, Harris MH, Li C, Lindsten T, Thompson CB (2005) Growth factor regulation of autophagy and cell survival in the absence of apoptosis. Cell 120:237–248

    Article  CAS  PubMed  Google Scholar 

  9. Morgan TM, Koreckij TD, Corey E (2009) Targeted therapy for advanced prostate cancer: inhibition of the PI3K/Akt/mTOR pathway. Curr Cancer Drug Targets 9:237–249

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Downward J (2004) PI 3-kinase, Akt and cell survival. Semin Cell Dev Biol 15:177–182

    Article  CAS  PubMed  Google Scholar 

  11. Liu P, Cheng H, Roberts TM, Zhao JJ (2009) Targeting the phosphoinositide 3-kinase pathway in cancer. Nat Rev Drug Discov 8:627–644

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Xu G, Zhang W, Bertram P, Zheng XF, McLeod H (2004) Pharmacogenomic profiling of the PI3K/PTEN-AKT-mTOR pathway in common human tumors. Int J Oncol 24:893–900

    CAS  PubMed  Google Scholar 

  13. Kim KW, Mutter RW, Cao C, Albert JM, Freeman M, Hallahan DE, Lu B (2006) Autophagy for cancer therapy through inhibition of pro-apoptotic proteins and mammalian target of rapamycin signaling. J Biol Chem 281:36883–36890

    Article  CAS  PubMed  Google Scholar 

  14. Sui YX, Li SG, Shi PY, Wu YJ, Li YX, Chen WY, Huang LY, Yao H, Lin X (2016) Ethyl acetate extract from Selaginella doederleinii Hieron inhibits the growth of human lung cancer cells A549 via caspase-dependent apoptosis pathway. J Ethnopharmacol 190:261–271

    Article  CAS  PubMed  Google Scholar 

  15. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  16. Chan LL, Shen D, Wilkinson AR, Patton W, Lai N, Chan E, Kuksin D, Lin B, Qiu J (2012) A novel image-based cytometry method for autophagy detection in living cells. Autophagy 8:1371–1382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Oeste CL, Seco E, Patton WF, Boya P, Perez-Sala D (2013) Interactions between autophagic and endo-lysosomal markers in endothelial cells. Histochem Cell Biol 139:659–670

    Article  CAS  PubMed  Google Scholar 

  18. Tanida I, Ueno T, Kominami E (2004) LC3 conjugation system in mammalian autophagy. Int J Biochem Cell Biol 36:2503–2518

    Article  CAS  PubMed  Google Scholar 

  19. Kabeya Y, Mizushima N, Yamamoto A, Oshitani-Okamoto S, Ohsumi Y, Yoshimori T (2004) LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci 117:2805–2812

    Article  CAS  PubMed  Google Scholar 

  20. Liang C, Feng P, Ku B, Dotan I, Canaani D, Oh BH (2006) Autophagic and tumour suppressor activity of a novel Beclin1-binding protein UVRAG. Nat Cell Biol 8:688–699

    Article  CAS  PubMed  Google Scholar 

  21. Wu J, Dang Y, Su W, Liu C, Ma H, Shan Y, Pei Y, Wan B, Guo J, Yu L (2006) Molecular cloning and characterization of rat LC3A and LC3B-- two novel markers of autophagosome. Biochem Bioph Res Commun 339:437–442

  22. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19:5720–5728

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Potze L, Mullauer FB, Colak S, Kessler JH, Medema JP (2014) Betulinic acid-induced mitochondria-dependent cell death is counterbalanced by an autophagic salvage response. Cell Death Dis 5:e1169

    Article  CAS  PubMed  Google Scholar 

  24. Blommaart EF, Luiken JJ, Blommaart PJ, van Woerkom GM, Meijer AJ (1995) Phosphorylation of ribosomal protein S6 is inhibitory for autophagy in isolated rat hepatocytes. J Biol Chem 270:2320–2326

    Article  CAS  PubMed  Google Scholar 

  25. Arico S, Petiot A, Bauvy C, Dubbelhuis PF, Meijer AJ, Codogno P (2001) The tumor suppressor PTEN positively regulates macroautophagy by inhibiting the phosphatidylinositol 3-kinase/protein kinase B pathway. J Biol Chem 276:35243–35246

    Article  CAS  PubMed  Google Scholar 

  26. Shigemitsu K, Tsujishita Y, Hara K, Nanahoshi M, Avruch J, Yonezawa K (1999) Regulation of translational effectors by amino acid and mammalian target of rapamycin signaling pathways. Possible involvement of autophagy in cultured hepatoma cells. J Biol Chem 274:1058–1065

    Article  CAS  PubMed  Google Scholar 

  27. Bursch W, Ellinger A, Gerner C, Frohwein U, Schulte-Hermann R (2000) Programmed cell death (PCD). Apoptosis, autophagic PCD, or others? Ann N Y Acad Sci 926:1–12

    Article  CAS  PubMed  Google Scholar 

  28. Corcelle EA, Puustinen P, Jaattele M (2009) Apoptosis and autophagy: targeting autophagy signaling in cancer cells-‘trick or treat’? FEBS J 276:6084–6096

    Article  CAS  PubMed  Google Scholar 

  29. Chiu W, Chen C-H, Chang J-N, Chen C-H, Hsu Y-H (2016) Far-infrared promotes burn wound healing by suppressing NLRP3 inflammasome caused by enhanced autophagy. J Mol Med 94:809–819

    Article  CAS  PubMed  Google Scholar 

  30. Suzuki SW, Yamamoto H, Oikawa Y, Kondo-Kakuta C, Kimura Y, Hirano H, Ohsumi Y (2015) Atg13 HORMA domain recruits Atg9 vesicles during autophagosome formation. PNAS 112:3350–3355

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Yang Y, Li Y, Chen X, Cheng X, Liao Y, Yu X (2016) Exosomal transfer of miR-30a between cardiomyocytes regulates autophagy after hypoxia. J Mol Med 94:711–724

    Article  CAS  PubMed  Google Scholar 

  32. Mochida K, Oikawa Y, Kimura Y, Kirisako H, Hirano H, Ohsumi Y, Nakatogawa H (2015) Receptor-mediated selective autophagy degrades the endoplasmic reticulum and the nucleus. Nature 522:359–362

    Article  CAS  PubMed  Google Scholar 

  33. Liu H, He Z, Simon H-U (2015) Protective role of autophagy and autophagy-related protein 5 in early tumorigenesis. J Mol Med 93:159–164

    Article  CAS  PubMed  Google Scholar 

  34. Yang Z, Goronzy JJ, Weyand CM (2015) Autophagy in autoimmune disease. J Mol Med 93:707–717

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G, Mukherjee C, Shi Y, Gélinas C, Fan Y et al (2006) Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell 10:51–64

  36. Chitra P, Saiprasad G, Manikandan R, Sudhandiran G (2015) Berberine inhibits Smad and non-Smad signaling cascades and enhances autophagy against pulmonary fibrosis. J Mol Med 93:1015–1031

    Article  CAS  PubMed  Google Scholar 

  37. Yu T, Zuber J, Li J (2015) Targeting autophagy in skin diseases. J Mol Med 93:31–38

    Article  PubMed  Google Scholar 

  38. Wang Y, Gao J, Zhang D, Zhang J, Ma J, Jiang H (2010) New insights into the antifibrotic effects of sorafenib on hepatic stellate cells and liver fibrosis. J Hepatol 53:132–144

    Article  CAS  PubMed  Google Scholar 

  39. Martelli AM, Evangelisti C, Chiarini F, Grimaldi C, Cappellini A, Ognibene A, McCubrey JA (2010) The emerging role of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin signaling network in normal myelopoiesis and leukemogenesis. Biochim Biophys Acta 1803:991–1002

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Nature Science Foundation of China (No. 21275028, 81303298, and 81202987), the Fujian Provincial Natural Science Foundation (No. 2016J01371), Program for New Century Excellent Talents in Fujian Province University (No.JA14128), Training project of young talents in health system of Fujian Province (No. 2016-ZQN-63), and the Fujian Agriculture and Forestry University Foundation for excellent youth teachers (No. xjq201414).

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou, 350108, China

    Yuxia Sui, Hong Yao, Shaoguang Li, Zhijun Li, Gang Wang, Shilan Lin, Youjia Wu, Yuxiang Li, Liying Huang & Xinhua Lin

  2. Department of Pharmacy, Provincial Clinical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, 350001, China

    Yuxia Sui

  3. Department of Pathology, Provincial Clinical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, 350001, China

    Long Jin

  4. Department of TCM Resource and Apitherapy, Bee Science College, Fujian Agriculture and Forestry University, Fuzhou, 350002, China

    Peiying Shi

  5. Institute of Nano Medicine Technology, Faculty of Pharmacy, Fujian Medical University, Fuzhou, 350108, China

    Qicai Liu

Authors
  1. Yuxia Sui
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hong Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shaoguang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Long Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peiying Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhijun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Shilan Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Youjia Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yuxiang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Liying Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Qicai Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Xinhua Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hong Yao, Qicai Liu or Xinhua Lin.

Ethics declarations

The animals were handled according to the guidelines of the China Animal Welfare Legislation, as provided by the Committee on Ethics in the Care and Use of Laboratory Animals of Fujian Medical University.

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Yuxia Sui and Hong Yao are contributed equally to this work.

Electronic supplementary material

ESM 1

(PDF 545 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sui, Y., Yao, H., Li, S. et al. Delicaflavone induces autophagic cell death in lung cancer via Akt/mTOR/p70S6K signaling pathway. J Mol Med 95, 311–322 (2017). https://doi.org/10.1007/s00109-016-1487-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00109-016-1487-z

Keyword

Search

Navigation