Molecular Medicine

, Volume 14, Issue 5–6, pp 264–275 | Cite as

Differential Expression of Receptor Tyrosine Kinases (RTKs) and IGF-I Pathway Activation in Human Uterine Leiomyomas

  • Linda Yu
  • Katrin Saile
  • Carol D. Swartz
  • Hong He
  • Xiaolin Zheng
  • Grace E. Kissling
  • Xudong Di
  • Shantelle Lucas
  • Stanley J. Robboy
  • Darlene Dixon
Research Article


Uterine leiomyomas (fibroids) are benign tumors that are prevalent in women of reproductive age. Research suggests that activated receptor tyrosine kinases (RTKs) play an important role in the enhanced proliferation observed in fibroids. In this study, a phospho-RTK array technique was used to detect RTK activity in leiomyomas compared with myometrial tissue. We found that fifteen out of seventeen RTKs evaluated in this study were highly expressed (P < 0.02–0.03) in the leiomyomas, and included the IGF-I/IGF-IR, EGF/EGFR, FGF/FGF-R, HGF/HGF-R, and PDGF/PDGF-R gene families. Due to the higher protein levels of IGF-IR observed in leiomyomas by us in earlier studies, we decided to focus on the activation of the IGF-IR, its downstream effectors, and MAPKp44/42 to confirm our earlier findings; and validate the significance of the increased IGF-IR phosphorylation observed by RTK array analysis in this study. We used immunolocalization, western blot, or immunoprecipitation studies and confirmed that leiomyomas overexpressed IGF-IRβ and phosphorylated IGF-IRβ. Additionally, we showed that the downstream effectors, Shc, Grb2, and MAPKp44/42 (P < 0.02–0.001) were also overexpressed and involved in IGF-IR signaling in these tumors, while IRS-I, PI3K, and AKT were not. In vitro studies showed that IGF-I (100 ng/mL) increased the proliferation of uterine leiomyoma cells (UtLM) (P < 0.0001), and that phosphorylated IGF-IRβ, Shc, and MAPKp44/42 were also overexpressed in IGF-I-treated UtLM cells (P < 0.05), similar to the tissue findings. A neutralizing antibody against the IGF-IRβ blocked these effects. These data indicate that overexpression of RTKs and, in particular, activation of the IGF-IR signaling pathway through Shc/Grb2/MAPK are important in mediating uterine leiomyoma growth. These data may provide new anti-tumor targets for noninvasive treatment of fibroids.



The authors would like to thank Dr. Gregg Richards for his extensive review of the original version of this manuscript. This research was supported, in part, by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences.


  1. 1.
    Payson M, Leppert P, Segars J. (2006) Epidemiology of myomas. Obstet. Gynecol. Clin. North Am. 33:1–11.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Mauskopf J, Flynn M, Thieda P, Spalding J, Duchane J. (2005) The economic impact of uterine fibroids in the United States: a summary of published estimates. J. Womens Health. (Larchmt) 14:692–703.CrossRefGoogle Scholar
  3. 3.
    Wallach EE, Vlahos NF. (2004) Uterine myomas: an overview of development, clinical features, and management. Obstet. Gynecol. 104:393–406.CrossRefPubMedGoogle Scholar
  4. 4.
    Bennasroune A, Gardin A, Aunis D, Cremel G, Hubert P. (2004) Tyrosine kinase receptors as attractive targets of cancer therapy. Crit. Rev. Oncol. Hematol. 50:23–38.CrossRefPubMedGoogle Scholar
  5. 5.
    Dixon D, He H, Haseman JK. (2000) Immunohistochemical localization of growth factors and their receptors in uterine leiomyomas and matched myometrium. Environ. Health Perspect. 108Suppl 5:795–802.CrossRefPubMedGoogle Scholar
  6. 6.
    Brahma PK, Martel KM, Christman GM. (2006) Future directions in myoma research. Obstet. Gynecol. Clin. North Am. 33:199–224, xiii.CrossRefPubMedGoogle Scholar
  7. 7.
    Walker CL, Stewart EA. (2005) Uterine fibroids: the elephant in the room. Science. 308:1589–92.CrossRefPubMedGoogle Scholar
  8. 8.
    Flake GP, Andersen J, Dixon D. (2003) Etiology and pathogenesis of uterine leiomyomas: a review. Environ. Health Perspect. 111:1037–54.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Burroughs KD, Howe SR, Okubo Y, Fuchs-Young R, LeRoith D, Walker CL. (2002) Dysregulation of IGF-I signaling in uterine leiomyoma. J. Endocrinol. 172:83–93.CrossRefPubMedGoogle Scholar
  10. 10.
    Wei J, Chiriboga L, Mizuguchi M, Yee H, Mittal K. (2005) Expression profile of tuberin and some potential tumorigenic factors in 60 patients with uterine leiomyomata. Mod. Pathol. 18:179–88.CrossRefPubMedGoogle Scholar
  11. 11.
    Giudice LC et al. (1993) Insulin-like growth factor (IGF), IGF binding protein (IGFBP), and IGF receptor gene expression and IGFBP synthesis in human uterine leiomyomata. Hum. Reprod. 8:1796–806.CrossRefPubMedGoogle Scholar
  12. 12.
    Rubin R, Baserga R. (1995) Insulin-like growth factor-I receptor. Its role in cell proliferation, apoptosis, and tumorigenicity. Lab. Invest. 73:311–31.PubMedGoogle Scholar
  13. 13.
    Werner H, Roberts CT Jr. (2003) The IGFI receptor gene: a molecular target for disrupted transcription factors. Genes Chromosomes Cancer 36:113–120.CrossRefPubMedGoogle Scholar
  14. 14.
    Mauro L, Surmacz E. (2004) IGF-I receptor, cell-cell adhesion, tumor development and progression. J. Mol. Histol. 35:247–53.CrossRefPubMedGoogle Scholar
  15. 15.
    Swartz CD, Afshari CA, Yu L, Hall KE, Dixon D. (2005) Estrogen-induced changes in IGF-I, Myb family and MAP kinase pathway genes in human uterine leiomyoma and normal uterine smooth muscle cell lines. Mol. Hum. Reprod. 11:441–50.CrossRefPubMedGoogle Scholar
  16. 16.
    Conover WJ, Iman RL. (1982) Analysis of covariance using the rank transformation. Biometrics. 38:715–24.CrossRefPubMedGoogle Scholar
  17. 17.
    Arslan AA et al. (2005) Gene expression studies provide clues to the pathogenesis of uterine leiomyoma: new evidence and a systematic review. Hum. Reprod. 20:852–63.CrossRefPubMedGoogle Scholar
  18. 18.
    Dixon D et al. (2002) Cell proliferation and apoptosis in human uterine leiomyomas and myometria. Virchows Arch. 441:53–62.CrossRefPubMedGoogle Scholar
  19. 19.
    Wang J et al. (2006) A novel selective progesterone receptor modulator asoprisnil (J867) down-regulates the expression of EGF, IGF-I, TGFbeta3 and their receptors in cultured uterine leiomyoma cells. Hum. Reprod. 21:1869–77.CrossRefPubMedGoogle Scholar
  20. 20.
    Harrison-Woolrych ML, Charnock-Jones DS, Smith SK. (1994) Quantification of messenger ribonucleic acid for epidermal growth factor in human myometrium and leiomyomata using reverse transcriptase polymerase chain reaction. J. Clin. Endocrinol. Metab. 78:1179–84.PubMedGoogle Scholar
  21. 21.
    Rein MS, Nowak RA. (1992) Biology of uterine myomas and myometrium in vitro. Semin. Reprod. Endocrinol. 10:310–9.CrossRefGoogle Scholar
  22. 22.
    Shimomura Y, Matsuo H, Samoto T, Maruo T. (1998) Up-regulation by progesterone of proliferating cell nuclear antigen and epidermal growth factor expression in human uterine leiomyoma. J. Clin. Endocrinol. Metab. 83:2192–8.PubMedGoogle Scholar
  23. 23.
    Stewart EA, Nowak RA. (1996) Leiomyomarelated bleeding: a classic hypothesis updated for the molecular era. Hum. Reprod. Update. 2:295–306.CrossRefPubMedGoogle Scholar
  24. 24.
    Mangrulkar RS, Ono M, Ishikawa M, Takashima S, Klagsbrun M, Nowak RA. (1995) Isolation and characterization of heparin-binding growth factors in human leiomyomas and normal myometrium. Biol. Reprod. 53:636–46.CrossRefPubMedGoogle Scholar
  25. 25.
    Wolanska M, Bankowski E. (2006) Fibroblast growth factors (FGF) in human myometrium and uterine leiomyomas in various stages of tumor growth. Biochimie. 88:141–6.CrossRefPubMedGoogle Scholar
  26. 26.
    Fayed YM, Tsibris JC, Langenberg PW, Robertson AL Jr. (1989) Human uterine leiomyoma cells: binding and growth responses to epidermal growth factor, platelet-derived growth factor, and insulin. Lab. Invest. 60:30–7.PubMedGoogle Scholar
  27. 27.
    Liang M, Wang H, Zhang Y, Lu S, Wang Z. (2006) Expression and functional analysis of platelet-derived growth factor in uterine leiomyomata. Cancer Biol. Ther. 5:28–33.CrossRefPubMedGoogle Scholar
  28. 28.
    Wolanska M, Bankowski E. (2007) Transforming growth factor beta and platelet-derived growth factor in human myometrium and in uterine leiomyomas at various stages of tumor growth. Eur. J. Obstet. Gynecol. Reprod. Biol. 130:238–44.CrossRefPubMedGoogle Scholar
  29. 29.
    Boehm KD, Daimon M, Gorodeski IG, Sheean LA, Utian WH, Ilan J. (1990) Expression of the insulin-like and platelet-derived growth factor genes in human uterine tissues. Mol. Reprod. Dev. 27:93–101.CrossRefPubMedGoogle Scholar
  30. 30.
    Hoppener JW et al. (1988) Expression of insulinlike growth factor-I and -II genes in human smooth muscle tumours. Embo. J. 7:1379–85.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Toscani GK et al. (2004) Gene expression and tyrosine kinase activity of insulin receptor in uterine leiomyoma and matched myometrium. Arch. Gynecol. Obstet. 270:170–3.CrossRefPubMedGoogle Scholar
  32. 32.
    Tommola P, Pekonen F, Rutanen EM. (1989) Binding of epidermal growth factor and insulin-like growth factor I in human myometrium and leiomyomata. Obstet. Gynecol. 74:658–62.PubMedGoogle Scholar
  33. 33.
    Chandrasekhar Y, Heiner J, Osuamkpe C, Nagamani M. (1992) Insulin-like growth factor I and II binding in human myometrium and leiomyomas. Am. J. Obstet. Gynecol. 166:64–9.CrossRefPubMedGoogle Scholar
  34. 34.
    Van der Ven LT et al. (1997) Expression of insulinlike growth factors (IGFs), their receptors and IGF binding protein-3 in normal, benign and malignant smooth muscle tissues. Br. J. Cancer. 75:1631–40.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Schlessinger J. (2000) Cell signaling by receptor tyrosine kinases. Cell. 103:211–25.CrossRefPubMedGoogle Scholar
  36. 36.
    Baserga R. (1998) The IGF-I Receptor in Normal and Abnormal Growth. In: Hormones and Growth Factors in Development and Neoplasia. Dickson RB, Salomon DS (ed.) Wiley-Liss Inc, Wilmington, DE, pp. 269–87.Google Scholar
  37. 37.
    van der Ven LT et al. (1994) Growth advantage of human leiomyoma cells compared to normal smooth-muscle cells due to enhanced sensitivity toward insulin-like growth factor I. Int. J. Cancer. 59:427–34.CrossRefPubMedGoogle Scholar
  38. 38.
    Englund K, Lindblom B, Carlstrom K, Gustavsson I, Sjoblom P, Blanck A. (2000) Gene expression and tissue concentrations of IGF-I in human myometrium and fibroids under different hormonal conditions. Mol. Hum. Reprod. 6:915–20.CrossRefPubMedGoogle Scholar
  39. 39.
    Wolanska M, Bankowski E. (2004) An accumulation of insulin-like growth factor I (IGF-I) in human myometrium and uterine leiomyomas in various stages of tumour growth. Eur. Cytokine Netw. 15:359–63.PubMedGoogle Scholar
  40. 40.
    O’Connor R. (2003) Regulation of IGF-I receptor signaling in tumor cells. Horm. Metab. Res. 35:771–7.CrossRefPubMedGoogle Scholar
  41. 41.
    Butler AA, Yakar S, Gewolb IH, Karas M, Okubo Y, LeRoith D. (1998) Insulin-like growth factor-I receptor signal transduction: at the interface between physiology and cell biology. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 121:19–26.CrossRefPubMedGoogle Scholar
  42. 42.
    Surmacz E. (2003) Growth factor receptors as therapeutic targets: strategies to inhibit the insulin-like growth factor I receptor. Oncogene. 22:6589–97.CrossRefPubMedGoogle Scholar
  43. 43.
    Kovacs KA et al. (2003) Differential expression of Akt/protein kinase B, Bcl-2 and Bax proteins in human leiomyoma and myometrium. J. Steroid Biochem. Mol. Biol. 87:233–40.CrossRefPubMedGoogle Scholar
  44. 44.
    Kovacs KA et al.(2007) Phosphorylation of PTEN (phosphatase and tensin homologue deleted on chromosome ten) protein is enhanced in human fibromyomatous uteri. J. Steroid Biochem. Mol. Biol. 103:196–9.CrossRefPubMedGoogle Scholar
  45. 45.
    Zumstein P, Stiles CD. (1987) Molecular cloning of gene sequences that are regulated by insulinlike growth factor I. J. Biol. Chem. 262:11252–60.PubMedGoogle Scholar
  46. 46.
    Strawn EY, Jr., Novy MJ, Burry KA, Bethea CL. (1995) Insulin-like growth factor I promotes leiomyoma cell growth in vitro. Am. J. Obstet. Gynecol. 172:1837–43; discussion 1843–4.CrossRefPubMedGoogle Scholar
  47. 47.
    Tao Y, Pinzi V, Bourhis J, Deutsch E. (2007) Mechanisms of disease: signaling of the insulin-like growth factor 1 receptor pathway—therapeutic perspectives in cancer. Nat. Clin. Pract. Oncol. 4:591–602.CrossRefPubMedGoogle Scholar
  48. 48.
    Clemmons DR. (2007) Modifying IGF1 activity: an approach to treat endocrine disorders, atherosclerosis and cancer. Nat. Rev. Drug Discov. 6:821–33.CrossRefPubMedGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2008

Authors and Affiliations

  • Linda Yu
    • 1
  • Katrin Saile
    • 1
  • Carol D. Swartz
    • 2
  • Hong He
    • 1
  • Xiaolin Zheng
    • 1
  • Grace E. Kissling
    • 3
  • Xudong Di
    • 1
  • Shantelle Lucas
    • 1
  • Stanley J. Robboy
    • 4
    • 5
  • Darlene Dixon
    • 1
  1. 1.Department of Health and Human Services (DHHS)Cellular and Molecular Pathology Branch, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health (NIH)Research Triangle ParkUSA
  2. 2.Environmental Carcinogenesis DivisionUS Environmental Protection AgencyResearch Triangle ParkUSA
  3. 3.Department of Health and Human Services (DHHS)Biostatistics Branch, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health (NIH)Research Triangle ParkUSA
  4. 4.Department of PathologyDuke University Medical CenterDurhamUSA
  5. 5.Department of Obstetrics and GynecologyDuke University Medical CenterDurhamUSA

Personalised recommendations