Skip to main content
SpringerLink
Log in

EGFR- and AKT-mediated reduction in PTEN expression contributes to tyrphostin resistance and is reversed by mTOR inhibition in endometrial cancer cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Loss or mutation of the PTEN (phosphatase and tensin homologue deleted on chromosome 10) gene is associated with resistance to epidermal growth factor receptor (EGFR) inhibitors. However, the mechanism underlying remains elusive. In this study, we aimed to explore whether sensitivity to the EGFR tyrosine kinase inhibitor (TKI) is affected by PTEN status in endometrial cancer cells. PTEN siRNA and the PTEN gene were transfected into HEC-1A and Ishikawa endometrial cancer cells using lentiviral vectors. Cells were treated under various concentrations of RG14620 and rapamycin, which are EGFR and mammalian target of rapamycin (mTOR) inhibitors, respectively. The IC50 of RG16420 was determined by using the MTT method. Cell apoptosis and the cell cycle were studied, and activation of EGFR, AKT, and p70S6 were detected by Western blot analysis. Loss of PTEN promoted cell proliferation and led to significant increases in the levels of EGFR, phospho-EGFR, AKT, phospho-AKT, and phospho-mTOR proteins. Ishikawa and HEC-1APTENkd cells that displayed loss and inactivation of PTEN function were resistant to RG14620. HEC-1A and IshikawaPTEN cells with intact PTEN were sensitive to RG14620. The combination of two inhibitors was more effective than both monotherapies, particularly in carcinoma cells with PTEN dysfunction. Decreased phospho-EGFR protein expression was observed in all cell lines that were sensitive to RG14620. Decreased phospho-AKT and phospho-p70S6 protein expression was observed in PTEN-intact cells that were sensitive to RG14620. PTEN loss results in resistance to EGFR TKI, which was reversed by PTEN reintroduction or mTOR inhibitor treatment. The combined treatment of EGFR TKI and the mTOR inhibitor provided a synergistic effect by promoting cell death in PTEN-deficient and PTEN-intact endometrial cancer cells, particularly in PTEN-deficient carcinoma cells with up-regulated EGFR activation.

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

Similar content being viewed by others

References

  1. Okuda T, Sekizawa A, Purwosunu Y, Nagatsuka M, Morioka M, Hayashi M, Okai T (2010) Genetics of endometrial cancers. Obstet Gynecol Int 2010:984013

    PubMed  Google Scholar 

  2. Doll A, Abal M, Rigau M, Monge M, Gonzalez M, Demajo S, Colas E, Llaurado M, Alazzouzi H, Planaguma J, Lohmann MA, Garcia J, Castellvi S, Ramon YCJ, Gil-Moreno A, Xercavins J, Alameda F, Reventos J (2008) Novel molecular profiles of endometrial cancer-new light through old windows. J Steroid Biochem Mol Biol 108:221–229

    Article  PubMed  CAS  Google Scholar 

  3. Herbst RS (2004) Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys 59:21–26

    Article  PubMed  CAS  Google Scholar 

  4. Laurent-Puig P, Lievre A, Blons H (2009) Mutations and response to epidermal growth factor receptor inhibitors. Clin Cancer Res 15:1133–1139

    Article  PubMed  CAS  Google Scholar 

  5. Chon HS, Hu W, Kavanagh JJ (2006) Targeted therapies in gynecologic cancers. Curr Cancer Drug Targets 6:333–363

    Article  PubMed  CAS  Google Scholar 

  6. Bansal N, Yendluri V, Wenham RM (2009) The molecular biology of endometrial cancers and the implications for pathogenesis, classification, and targeted therapies. Cancer Control 16:8–13

    PubMed  Google Scholar 

  7. Cheung AN (2007) Molecular targets in gynaecological cancers. Pathology 39:26–45

    Article  PubMed  CAS  Google Scholar 

  8. Gadducci A, Tana R, Cosio S, Fanucchi A, Genazzani AR (2008) Molecular target therapies in endometrial cancer: from the basic research to the clinic. Gynecol Endocrinol 24:239–249

    Article  PubMed  CAS  Google Scholar 

  9. She QB, Solit D, Basso A, Moasser MM (2003) Resistance to gefitinib in PTEN-null HER-overexpressing tumor cells can be overcome through restoration of PTEN function or pharmacologic modulation of constitutive phosphatidylinositol 3′-kinase/AKT pathway signaling. Clin Cancer Res 9:4340–4346

    PubMed  CAS  Google Scholar 

  10. Festuccia C, Muzi P, Millimaggi D, Biordi L, Gravina GL, Speca S, Angelucci A, Dolo V, Vicentini C, Bologna M (2005) Molecular aspects of gefitinib antiproliferative and pro-apoptotic effects in PTEN-positive and PTEN-negative prostate cancer cell lines. Endocr Relat Cancer 12:983–998

    Article  PubMed  CAS  Google Scholar 

  11. Mutter GL (2001) PTEN, a protean tumor suppressor. Am J Pathol 158:1895–1898

    Article  PubMed  CAS  Google Scholar 

  12. Mutter GL, Lin MC, Fitzgerald JT, Kum JB, Baak JP, Lees JA, Weng LP, Eng C (2000) Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. J Natl Cancer Inst 92:924–930

    Article  PubMed  CAS  Google Scholar 

  13. Uegaki K, Kanamori Y, Kigawa J, Kawaguchi W, Kaneko R, Naniwa J, Takahashi M, Shimada M, Oishi T, Itamochi H, Terakawa N (2006) PTEN is involved in the signal transduction pathway of contact inhibition in endometrial cells. Cell Tissue Res 323:523–528

    Article  PubMed  CAS  Google Scholar 

  14. DeGraffenried LA, Fulcher L, Friedrichs WE, Grunwald V, Ray RB, Hidalgo M (2004) Reduced PTEN expression in breast cancer cells confers susceptibility to inhibitors of the PI3 kinase/AKT pathway. Ann Oncol 15:1510–1516

    Article  PubMed  CAS  Google Scholar 

  15. Neshat MS, Mellinghoff IK, Tran C, Stiles B, Thomas G, Petersen R, Frost P, Gibbons JJ, Wu H, Sawyers CL (2001) Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci USA 98:10314–10319

    Article  PubMed  CAS  Google Scholar 

  16. Albitar L, Carter MB, Davies S, Leslie KK (2007) Consequences of the loss of p53, RB1, and PTEN: relationship to gefitinib resistance in endometrial cancer. Gynecol Oncol 106:94–104

    Article  PubMed  CAS  Google Scholar 

  17. Chon HS, Hu W, Kavanagh JJ (2006) Targeted therapies in gynecologic cancers. Curr Cancer Drug Targets 6:333–363

    Article  PubMed  CAS  Google Scholar 

  18. Wang MY, Lu KV, Zhu S, Dia EQ, Vivanco I, Shackleford GM, Cavenee WK, Mellinghoff IK, Cloughesy TF, Sawyers CL, Mischel PS (2006) Mammalian target of rapamycin inhibition promotes response to epidermal growth factor receptor kinase inhibitors in PTEN-deficient and PTEN-intact glioblastoma cells. Cancer Res 66:7864–7869

    Article  PubMed  CAS  Google Scholar 

  19. Oza AM, Eisenhauer EA, Elit L, Cutz JC, Sakurada A, Tsao MS, Hoskins PJ, Biagi J, Ghatage P, Mazurka J, Provencher D, Dore N, Dancey J, Fyles A (2008) Phase II study of erlotinib in recurrent or metastatic endometrial cancer: NCIC IND-148. J Clin Oncol 26:4319–4325

    Article  PubMed  CAS  Google Scholar 

  20. Tang LL, Yokoyama Y, Wan X, Iwagaki S, Niwa K, Tamaya T (2006) PTEN sensitizes epidermal growth factor-mediated proliferation in endometrial carcinoma cells. Oncol Rep 15:855–859

    PubMed  CAS  Google Scholar 

  21. Xiao L, Yang YB, Li XM, Xu CF, Li T, Wang XY (2010) Differential sensitivity of human endometrial carcinoma cells with different PTEN expression to mitogen-activated protein kinase signaling inhibits and implications for therapy. J Cancer Res Clin Oncol 136:1089–1099

    Article  PubMed  CAS  Google Scholar 

  22. Xiao L, Yang YB, Shen HM, Xu CF, Wang XY, Li XM (2009) Effect of signal transduction inhibitors on human endometrial carcinoma cells with differential PTEN gene expression. Zhonghua Fu Chan Ke Za Zhi 44:681–685

    PubMed  CAS  Google Scholar 

  23. Wan X, Yokoyama Y, Shinohara A, Takahashi Y, Tamaya T (2002) PTEN augments staurosporine-induced apoptosis in PTEN-null Ishikawa cells by downregulating PI3K/AKT signaling pathway. Cell Death Differ 9:414–420

    Article  PubMed  CAS  Google Scholar 

  24. Wan X, Li J, Xie X, Lu W (2007) PTEN augments doxorubicin-induced apoptosis in PTEN-null Ishikawa cells. Int J Gynecol Cancer 17:808–812

    Article  PubMed  CAS  Google Scholar 

  25. Zhao LJ, Liu N, Li XP, Wang JL, Wei LH (2010) Phosphatase and tensin homolog gene inhibits the effect induced by gonadotropin-releasing hormone subtypes in human endometrial carcinoma cells. Chin Med J (Engl) 123:1170–1175

    CAS  Google Scholar 

  26. Sakurada A, Hamada H, Fukushige S, Yokoyama T, Yoshinaga K, Furukawa T, Sato S, Yajima A, Sato M, Fujimura S, Horii A (1999) Adenovirus-mediated delivery of the PTEN gene inhibits cell growth by induction of apoptosis in endometrial cancer. Int J Oncol 15:1069–1074

    PubMed  CAS  Google Scholar 

  27. Wendel HG, Malina A, Zhao Z, Zender L, Kogan SC, Cordon-Cardo C, Pelletier J, Lowe SW (2006) Determinants of sensitivity and resistance to rapamycin-chemotherapy drug combinations in vivo. Cancer Res 66:7639–7646

    Article  PubMed  CAS  Google Scholar 

  28. Neshat MS, Mellinghoff IK, Tran C, Stiles B, Thomas G, Petersen R, Frost P, Gibbons JJ, Wu H, Sawyers CL (2001) Enhanced sensitivity of PTEN-deficient tumors to inhibition of FRAP/mTOR. Proc Natl Acad Sci USA 98:10314–10319

    Article  PubMed  CAS  Google Scholar 

  29. Darb-Esfahani S, Faggad A, Noske A, Weichert W, Buckendahl AC, Muller B, Budczies J, Roske A, Dietel M, Denkert C (2009) Phospho-mTOR and phospho-4EBP1 in endometrial adenocarcinoma: association with stage and grade in vivo and link with response to rapamycin treatment in vitro. J Cancer Res Clin Oncol 135:933–941

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by a grant from the Science and Technology Development Fund of the Macao Special Administrative Region (002/2009/A) and the National Natural Science Foundation of China (30772332). We thank Professor Zehua Wang (Department of Obstetrics & Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, P.R. China) for supplying the cell lines.

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, The Third Affiliated Hospital, Sun Yat-sen University, 600 TiaHe Road, Guangzhou, 510630, People’s Republic of China

    Tian Li, Yuebo Yang, Xiaomao Li & Chengfang Xu

  2. The School of Health Sciences, Macao Polytechnic Institute, 7th Floor, Meng Tak building, Rua de Luis Gonzaga Gomes, Macao, People’s Republic of China

    Lirong Meng

Authors
  1. Tian Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuebo Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaomao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chengfang Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lirong Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lirong Meng.

Additional information

Tian Li and Yuebo Yang contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, T., Yang, Y., Li, X. et al. EGFR- and AKT-mediated reduction in PTEN expression contributes to tyrphostin resistance and is reversed by mTOR inhibition in endometrial cancer cells. Mol Cell Biochem 361, 19–29 (2012). https://doi.org/10.1007/s11010-011-1082-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-011-1082-0

Keywords

Search

Navigation