Original Article | Published:

A regulatory circuit composed of DNA methyltransferases and receptor tyrosine kinases controls lung cancer cell aggressiveness

Oncogene volume 36, pages 69196928 (14 December 2017) | Download Citation

Abstract

Overexpression of DNMT1 and KIT is prevalent in lung cancer, yet the underlying molecular mechanisms are poorly understood. While the deregulated activation of DNMT1 or KIT has been implicated in lung cancer pathogenesis, whether and how DNMT1 and KIT orchestrate lung tumorigenesis are unclear. Here, using human lung cancer tissue microarrays and fresh frozen tissues, we found that the overexpression of DNMT1 is positively correlated with the upregulation of KIT in tumor tissues. We demonstrated that DNMT1 and KIT form a positive regulatory loop, in which ectopic DNMT1 expression increases, whereas targeted DNMT1 depletion abrogates KIT signaling cascade through Sp1/miR-29b network. Conversely, an increase of KIT levels augments, but a reduction of KIT expression ablates DNMT1 transcription by STAT3 pathway leading to in-parallel modification of the DNA methylation profiles. We provided evidence that KIT inactivation induces global DNA hypomethylation, restores the expression of tumor suppressor p15INK4B through promoter demethylation; in turn, DNMT1 dysfunction impairs KIT kinase signaling. Functionally, KIT and DNMT1 co-expression promotes, whereas dual inactivation of them suppresses, lung cancer cell proliferation and metastatic growth in vitro and in vivo, in a synergistic manner. These findings demonstrate the regulatory and functional interplay between DNA methylation and tyrosine kinase signaling in propelling tumorigenesis, providing a widely applicable approach for targeting lung cancer.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , , . DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 1999; 99: 247–257.

  2. 2.

    , , , , , . De novo methylation of nucleosomal DNA by the mammalian Dnmt1 and Dnmt3A DNA methyltransferases. Biochemistry 2005; 44: 9899–9904.

  3. 3.

    , , , , , et al. Global analysis of DNA methylation by methyl-capture sequencing reveals epigenetic control of cisplatin resistance in ovarian cancer cell. PloS one 2011; 6: e29450.

  4. 4.

    , , , , , et al. In vivo control of CpG and non-CpG DNA methylation by DNA methyltransferases. PLoS genet 2012; 8: e1002750.

  5. 5.

    , . New concepts in DNA methylation. Trends biochem sci 2014; 39: 310–318.

  6. 6.

    , , , , , et al. Phosphorylation of TET proteins is regulated via O-GlcNAcylation by the glycosyltransferase OGT. J Biol Chem 2015; 290: 4801–4812.

  7. 7.

    , , , , , . Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIH 3T3 cells. Proc Natl Acad Sci USA 1993; 90: 8891–8895.

  8. 8.

    , , , , , et al. MicroRNA -29b induces global DNA hypomethylation and tumor suppressor gene re-expression in acute myeloid leukemia by targeting directly DNMT3A and 3B and indirectly DNMT1. Blood 2009; 113: 6411–6418.

  9. 9.

    , , , , , et al. Bortezomib induces DNA hypomethylation and silenced gene transcription by interfering with Sp1/NF-kappaB-dependent DNA methyltransferase activity in acute myeloid leukemia. Blood 2008; 111: 2364–2373.

  10. 10.

    , , , , , et al. A phase II study of dasatinib in patients with chemosensitive relapsed small cell lung cancer (Cancer and Leukemia Group B 30602). J thorac oncol 2010; 5: 380–384.

  11. 11.

    , , , , , et al. MicroRNA-29 family reverts aberrant methylation in lung cancer by targeting DNA methyltransferases 3A and 3B. Proc Natl Acad Sci USA 2007; 104: 15805–15810.

  12. 12.

    , , , , , . Alteration of DNA methyltransferases contributes to 5'CpG methylation and poor prognosis in lung cancer. Lung Cancer 2007; 55: 205–213.

  13. 13.

    , , , , , et al. Detection of overexpressed and phosphorylated wild-type kit receptor in surgical specimens of small cell lung cancer. Clin Cancer Res 2004; 10: 8214–8219.

  14. 14.

    , . Imatinib inhibits c-Kit-induced hypoxia-inducible factor-1alpha activity and vascular endothelial growth factor expression in small cell lung cancer cells. Mol cancer therapeut 2006; 5: 1415–1422.

  15. 15.

    , , . Regulation of DNA methyltransferase 1 by the pRb/E2F1 pathway. Cancer Res 2005; 65: 3624–3632.

  16. 16.

    , , , , . Transcription of mouse DNA methyltransferase 1 (Dnmt1) is regulated by both E2F-Rb-HDAC-dependent and -independent pathways. Nucleic Acids Res 2003; 31: 3101–3113.

  17. 17.

    , , , . Mechanisms of metastasis as related to receptor tyrosine kinases in small-cell lung cancer. J environ pathol toxicol oncol 2003; 22: 147–165.

  18. 18.

    , , , , , et al. Elimination of human lung cancer stem cells through targeting of the stem cell factor-c-kit autocrine signaling loop. Cancer Res 2010; 70: 338–346.

  19. 19.

    , , , , , et al. Preferential localization of c-kit product in tissue mast cells, basal cells of skin, epithelial cells of breast, small cell lung carcinoma and seminoma/dysgerminoma in human: immunohistochemical study on formalin-fixed, paraffin-embedded tissues. Virchows Archiv 1994; 424: 135–141.

  20. 20.

    , , , , , et al. Expression and mutational status of c-kit in small-cell lung cancer: prognostic relevance. Clin Cancer Res 2004; 10: 4101–4108.

  21. 21.

    , , , , , . Cancer statistics, 2007. CA: cancer j clin 2007; 57: 43–66.

  22. 22.

    , . Receptor tyrosine kinases as targets for anticancer therapeutics. Curr Med Chem 2005; 12: 1773–1781.

  23. 23.

    , , , , , et al. Recent developments of targeted therapies in the treatment of non-small cell lung cancer. Curr Drug Discov Technol 2009; 6: 91–102.

  24. 24.

    , , , . Role of tyrosine kinase inhibitors in lung cancer. Anticancer Agents Med Chem 2009; 9: 569–575.

  25. 25.

    , , , , , et al. Phase I study of PKC412 (N-benzoyl-staurosporine), a novel oral protein kinase C inhibitor, combined with gemcitabine and cisplatin in patients with non-small-cell lung cancer. Ann Oncol 2004; 15: 316–323.

  26. 26.

    , , , , , et al. Phase I study of decitabine-mediated gene expression in patients with cancers involving the lungs, esophagus, or pleura. Clin Cancer Res 2006; 12: 5777–5785.

  27. 27.

    , , , , , et al. Phase-I study of sagopilone in combination with cisplatin in chemotherapy-naive patients with metastasised small-cell lung cancer. Eur J Cancer 2013; 49: 2461–2468.

  28. 28.

    , , , , , et al. A Cullin3-KLHL20 ubiquitin ligase-dependent pathway targets PML to potentiate HIF-1 signaling and prostate cancer progression. Cancer Cell 2011; 20: 214–228.

  29. 29.

    , , , , , . The DNA methyltransferase DNMT1 and tyrosine-protein kinase KIT cooperatively promote resistance to 5-Aza-2'-deoxycytidine (decitabine) and midostaurin (PKC412) in lung cancer cells. J Biol Chem 2015; 290: 18480–18494.

  30. 30.

    , , , , , . Transcriptional suppression of mir-29b-1/mir-29a promoter by c-Myc, hedgehog, and NF-kappaB. J cell biochem 2010; 110: 1155–1164.

  31. 31.

    , , , , , et al. Sp1/NFkappaB/HDAC/miR-29b regulatory network in KIT-driven myeloid leukemia. Cancer Cell 2010; 17: 333–347.

  32. 32.

    , . Cisplatin regulates the MAPK kinase pathway to induce increased expression of DNA repair gene ERCC1 and increase melanoma chemoresistance. Oncogene 2012; 31: 2412–2422.

  33. 33.

    , , , , . Increasing chemotherapy in small-cell lung cancer: from dose intensity and density to megadoses. oncologist 2007; 12: 79–89.

  34. 34.

    , , , , . Targeted therapies in small cell lung cancer: a review. Therapeut adv med oncol 2010; 2: 25–37.

  35. 35.

    , , , , , et al. DNA methylation in small cell lung cancer defines distinct disease subtypes and correlates with high expression of EZH2. Oncogene 2015; 34: 5869–5878.

  36. 36.

    , , , , , et al. Dysregulation of p53/Sp1 control leads to DNA methyltransferase-1 overexpression in lung cancer. Cancer Res 2010; 70: 5807–5817.

  37. 37.

    , , , , , et al. Checkpoint kinase inhibitor AZD7762 overcomes cisplatin resistance in clear cell carcinoma of the ovary. Int j gynecol cancer 2014; 24: 61–69.

  38. 38.

    , , , , , . AKT1 amplification regulates cisplatin resistance in human lung cancer cells through the mammalian target of rapamycin/p70S6K1 pathway. Cancer Res 2007; 67: 6325–6332.

  39. 39.

    , , , , , et al. Phase II trial of imatinib maintenance therapy after irinotecan and cisplatin in patients with c-Kit-positive, extensive-stage small-cell lung cancer. Clin lung cancer 2010; 11: 223–227.

  40. 40.

    , , , , , et al. Fatty acid-binding protein FABP4 mechanistically links obesity with aggressive AML by enhancing aberrant DNA methylation in AML cells. Leukemia 2016; 31: 1434–1442.

  41. 41.

    , , , , , et al. Restoration of miR-101 suppresses lung tumorigenesis through inhibition of DNMT3a-dependent DNA methylation. Cell death dis 2014; 5: e1413.

  42. 42.

    , , , , , et al. AML1/ETO cooperates with HIF1alpha to promote leukemogenesis through DNMT3a transactivation. Leukemia 2015; 29: 1730–1740.

Download references

Acknowledgements

This work was supported in part by Hormel Foundation (S. Liu) and National Cancer Institute (Bethesda, MD, USA) grants R01CA149623 (S. Liu) and R21CA155915 (S. Liu).

Author contributions

SL designed the research; FY, NS, JP, NZ and BD performed research; YY, PY and JRM provided the patient samples; FY, BL, PY, JRM and SL analyzed data and wrote the paper; SL conceived the idea and supervised the whole project. All authors discussed the results and commented on the manuscript.

Author information

Author notes

    • F Yan
    •  & N Shen

    These authors contributed equally to this work.

Affiliations

  1. The Hormel Institute, University of Minnesota, Austin, MN, USA

    • F Yan
    • , N Shen
    • , J Pang
    • , N Zhao
    •  & S Liu
  2. Division of Epidemiology, Mayo Clinic, Rochester, MN, USA

    • B Deng
    • , Y Yang
    •  & P Yang
  3. Department of Microbiology and Immunology, University of Louisville, Louisville, KY, USA

    • B Li
  4. Department of Medical Oncology, Mayo Clinic, Rochester, MN, USA

    • J R Molina

Authors

  1. Search for F Yan in:

  2. Search for N Shen in:

  3. Search for J Pang in:

  4. Search for N Zhao in:

  5. Search for B Deng in:

  6. Search for B Li in:

  7. Search for Y Yang in:

  8. Search for P Yang in:

  9. Search for J R Molina in:

  10. Search for S Liu in:

Competing interests

The authors declare no conflict of interest.

Corresponding author

Correspondence to S Liu.

Supplementary information

Word documents

  1. 1.

    Supplementary Materials

About this article

Publication history

Received

Revised

Accepted

Published

DOI

https://doi.org/10.1038/onc.2017.305

Supplementary Information accompanies this paper on the Oncogene website (http://www.nature.com/onc)

Further reading