Molecular Genetics and Genomics

, Volume 294, Issue 1, pp 85–93 | Cite as

HOXA4-regulated miR-138 suppresses proliferation and gefitinib resistance in non-small cell lung cancer

  • Xiaomei Tang
  • Jiying Jiang
  • Jinbao Zhu
  • Nan He
  • Jinlong TanEmail author
Original Article


Many non-small cell lung cancer (NSCLC) patients initially benefiting from gefitinib are confronted with acquired resistance. MiR-138 was previously stated as a growth inhibitor of several cancer cell lines including NSCLC cells and its expression level was significantly lower in gefitinib-resistant cells. The role of miR-138 in NSCLC cell lines PC9 and A549 was verified using methyl thiazolyl tetrazolium (MTT) assay and colony formation assay. Quantitative real-time PCR (RT-PCR) was employed to assess the level of miR-138 in gefitinib-sensitive PC9 cells and gefitinib-resistant PC9GR cells. Bioinformatic algorithms (TargetScan) and rVISTA 2.0 were used to predict binding sites on miR-138 and its target genes. MiR-138 inhibited cell proliferation of PC9 and A549 cells. In PC9GR cells, miR-138 expression was inhibited. Gefitinib treatment negatively regulated miR-138 in PC9 cells. Transfection of PC9GR cells with miR-138 mimics significantly reduced cell viability. MiR-138 was directly regulated by Homeobox A4 (HOXA4) via an HOXA4-binding site on the promoter region. TargetScan predicted numerous miR-138 target genes and EGFR was found to be the functional downstream effector of miR-138. We demonstrated that miR-138 is regulated by HOXA4 and exerts its functions via inhibiting EGFR expression in NSCLC cells.


Non-small cell lung cancer (NSCLC) Gefitinib MiR-138 Homeobox A4 (HOXA4) Epidermal growth factor receptor (EGFR) 




Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Not applicable.


  1. Agarwal V, Bell GW, Nam JW, Bartel DP (2015) Predicting effective microRNA target sites in mammalian mRNAs. Elife. Google Scholar
  2. Chen D, Guo W, Qiu Z, Wang Q, Li Y, Liang L, Liu L, Huang S, Zhao Y, He X (2015) MicroRNA-30d-5p inhibits tumour cell proliferation and motility by directly targeting CCNE2 in non-small cell lung cancer. Cancer Lett 362(2):208–217CrossRefGoogle Scholar
  3. Ercan D, Zejnullahu K, Yonesaka K, Xiao Y, Capelletti M, Rogers A, Lifshits E, Brown A, Lee C, Christensen JG, Kwiatkowski DJ, Engelman JA, Janne PA (2010) Amplification of EGFR T790M causes resistance to an irreversible EGFR inhibitor. Oncogene 29(16):2346–2356CrossRefGoogle Scholar
  4. Fenizia F, De Luca A, Pasquale R, Sacco A, Forgione L, Lambiase M, Iannaccone A, Chicchinelli N, Franco R, Rossi A, Morabito A, Rocco G, Piccirillo MC, Normanno N (2015) EGFR mutations in lung cancer: from tissue testing to liquid biopsy. Future Oncol 11(11):1611–1623CrossRefGoogle Scholar
  5. Fortunato O, Boeri M, Verri C, Moro M, Sozzi G (2014) Therapeutic use of microRNAs in lung cancer. BioMed Res Int 2014:756975CrossRefGoogle Scholar
  6. Gao Y, Fan X, Li W, Ping W, Deng Y, Fu X (2014) miR-138-5p reverses gefitinib resistance in non-small cell lung cancer cells via negatively regulating G protein-coupled receptor 124. Biochem Biophys Res Commun 446(1):179–186CrossRefGoogle Scholar
  7. Inamura K, Ninomiya H, Ishikawa Y, Matsubara O (2010) Is the epidermal growth factor receptor status in lung cancers reflected in clinicopathologic features? Arch Pathol Lab Med 134(1):66–72Google Scholar
  8. Inoue A, Yoshida K, Morita S, Imamura F, Seto T, Okamoto I, Nakagawa K, Yamamoto N, Muto S, Fukuoka M (2016) Characteristics and overall survival of EGFR mutation-positive non-small cell lung cancer treated with EGFR tyrosine kinase inhibitors: a retrospective analysis for 1660 Japanese patients. Jpn J Clin Oncol 46(5):462–467CrossRefGoogle Scholar
  9. Iorio MV, Ferracin M, Liu CG, Veronese A, Spizzo R, Sabbioni S, Magri E, Pedriali M, Fabbri M, Campiglio M, Menard S, Palazzo JP, Rosenberg A, Musiani P, Volinia S, Nenci I, Calin GA, Querzoli P, Negrini M, Croce CM (2005) MicroRNA gene expression deregulation in human breast cancer. Cancer Res 65(16):7065–7070CrossRefGoogle Scholar
  10. Kasinski AL, Slack FJ (2011) Epigenetics and genetics. MicroRNAs en route to the clinic: progress in validating and targeting microRNAs for cancer therapy. Nat Rev Cancer 11(12):849–864CrossRefGoogle Scholar
  11. Kasinski AL, Kelnar K, Stahlhut C, Orellana E, Zhao J, Shimer E, Dysart S, Chen X, Bader AG, Slack FJ (2015) A combinatorial microRNA therapeutics approach to suppressing non-small cell lung cancer. Oncogene 34(27):3547–3555CrossRefGoogle Scholar
  12. Klausen C, Leung PC, Auersperg N (2009) Cell motility and spreading are suppressed by HOXA4 in ovarian cancer cells: possible involvement of beta1 integrin. Mol Cancer Res 7(9):1425–1437CrossRefGoogle Scholar
  13. Kobayashi S, Boggon TJ, Dayaram T, Janne PA, Kocher O, Meyerson M, Johnson BE, Eck MJ, Tenen DG, Halmos B (2005) EGFR mutation and resistance of non-small-cell lung cancer to gefitinib. N Engl J Med 352(8):786–792CrossRefGoogle Scholar
  14. Li J, Wang Q, Wen R, Liang J, Zhong X, Yang W, Su D, Tang J (2015) MiR-138 inhibits cell proliferation and reverses epithelial-mesenchymal transition in non-small cell lung cancer cells by targeting GIT1 and SEMA4C. J Cell Mol Med 19(12):2793–2805CrossRefGoogle Scholar
  15. Lin JJ, Cardarella S, Lydon CA, Dahlberg SE, Jackman DM, Janne PA, Johnson BE (2016) Five-year survival in EGFR-mutant metastatic lung adenocarcinoma treated with EGFR-TKIs. J Thorac Oncol 11(4):556–565CrossRefGoogle Scholar
  16. Liu X, Chen Z, Yu J, Xia J, Zhou X (2009a) MicroRNA profiling and head and neck cancer. Comp Funct Genom 2009(837514):11Google Scholar
  17. Liu X, Jiang L, Wang A, Yu J, Shi F, Zhou X (2009b) MicroRNA-138 suppresses invasion and promotes apoptosis in head and neck squamous cell carcinoma cell lines. Cancer Lett 286(2):217–222CrossRefGoogle Scholar
  18. Loots GG, Ovcharenko I (2004) rVISTA 2.0: evolutionary analysis of transcription factor binding sites. Nucleic Acids Res 32(web server issue):W217–W221CrossRefGoogle Scholar
  19. Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, Downing JR, Jacks T, Horvitz HR, Golub TR (2005) MicroRNA expression profiles classify human cancers. Nature 435(7043):834–838CrossRefGoogle Scholar
  20. Macfarlane LA, Murphy PR (2010) MicroRNA: biogenesis, function and role in cancer. Curr Genom 11(7):537–561CrossRefGoogle Scholar
  21. Miller KR, Patel JN, Zhang Q, Norris EJ, Symanowski J, Michener C, Sehouli J, Braicu I, Destephanis DD, Sutker AP (2018) HOXA4/HOXB3 gene expression signature as a biomarker of recurrence in patients with high-grade serous ovarian cancer following primary cytoreductive surgery and first-line adjuvant chemotherapy. Gynecol Oncol 149(1):155–162CrossRefGoogle Scholar
  22. Ota T, Klausen C, Salamanca MC, Woo HL, Leung PC, Auersperg N (2009) Expression and function of HOXA genes in normal and neoplastic ovarian epithelial cells. Differentiation 77(2):162–171CrossRefGoogle Scholar
  23. Scagliotti GV, Selvaggi G, Novello S, Hirsch FR (2004) The biology of epidermal growth factor receptor in lung cancer. Clin Cancer Res 10(12 Pt 2):4227s–4232sCrossRefGoogle Scholar
  24. Siegel RL, Miller KD, Jemal A (2015) Cancer statistics, 2015. CA Cancer J Clin 65(1):5–29CrossRefGoogle Scholar
  25. Stinchcombe TE (2014) Recent advances in the treatment of non-small cell and small cell lung cancer. F1000Prime Rep 6:117CrossRefGoogle Scholar
  26. Sun C, Li S, Yang C, Xi Y, Wang L, Zhang F, Li D (2016) MicroRNA-187-3p mitigates non-small cell lung cancer (NSCLC) development through down-regulation of BCL6. Biochem Biophys Res Commun 471(1):82–88CrossRefGoogle Scholar
  27. Uzel EK, Abacioglu U (2015) Treatment of early stage non-small cell lung cancer: surgery or stereotactic ablative radiotherapy? Balkan Med J 32(1):8–16Google Scholar
  28. van Zandwijk N, Fong KM (2015) Update in lung cancer: prologue to a modern review series. Respirology 20(2):183–184CrossRefGoogle Scholar
  29. Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, Petrocca F, Visone R, Iorio M, Roldo C, Ferracin M, Prueitt RL, Yanaihara N, Lanza G, Scarpa A, Vecchione A, Negrini M, Harris CC, Croce CM (2006) A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci USA 103(7):2257–2261CrossRefGoogle Scholar
  30. Xu C, Zheng Y, Lian D, Ye S, Yang J, Zeng Z (2015) Analysis of microRNA expression profile identifies novel biomarkers for non-small cell lung cancer. Tumori 101(1):104–110CrossRefGoogle Scholar
  31. Ye XW, Yu H, Jin YK, Jing XT, Xu M, Wan ZF, Zhang XY (2015) miR-138 inhibits proliferation by targeting 3-phosphoinositide-dependent protein kinase-1 in non-small cell lung cancer cells. Clin Respir J 9(1):27–33CrossRefGoogle Scholar
  32. Yu T, Li J, Yan M, Liu L, Lin H, Zhao F, Sun L, Zhang Y, Cui Y, Zhang F, Li J, He X, Yao M (2015) MicroRNA-193a-3p and -5p suppress the metastasis of human non-small-cell lung cancer by downregulating the ERBB4/PIK3R3/mTOR/S6K2 signaling pathway. Oncogene 34(4):413–423CrossRefGoogle Scholar
  33. Yun CH, Mengwasser KE, Toms AV, Woo MS, Greulich H, Wong KK, Meyerson M, Eck MJ (2008) The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP. Proc Natl Acad Sci USA 105(6):2070–2075CrossRefGoogle Scholar
  34. Zappa C, Mousa SA (2016) Non-small cell lung cancer: current treatment and future advances. Transl Lung Cancer Res 5(3):288–300CrossRefGoogle Scholar
  35. Zhang H, Zhang H, Zhao M, Lv Z, Zhang X, Qin X, Wang H, Wang S, Su J, Lv X (2013) MiR-138 inhibits tumor growth through repression of EZH2 in non-small cell lung cancer. Cell Physiol Biochem 31(1):56–65CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Xiaomei Tang
    • 1
  • Jiying Jiang
    • 2
  • Jinbao Zhu
    • 3
  • Nan He
    • 4
  • Jinlong Tan
    • 5
    Email author
  1. 1.Jiangxi Chest HospitalNanchangChina
  2. 2.Central Hospital of Wannian CountyWannian CountyChina
  3. 3.Yugan County People’s HospitalYuganChina
  4. 4.Guangdong Ascendas Genomics Technology Co., Ltd.ZhongshanChina
  5. 5.ShangRao People Hospital of JiangxiShangraoChina

Personalised recommendations