Download PDF
Horm Metab Res 2003; 35(11/12): 850-856
DOI: 10.1055/s-2004-814144
Original
© Georg Thieme Verlag Stuttgart · New York

Inhibition of the Biologic Response to Insulin-like Growth Factor I in MCF-7 Breast Cancer Cells by a New Monoclonal Antibody to the Insulin-like Growth Factor-I Receptor. The Importance of Receptor Down-regulation

P.-G.  Jackson-Booth1 , C.  Terry1 , B.  Lackey1 , M.  Lopaczynska1 , P.  Nissley1
  • 1Metabolism Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892 USA
Further Information

Publication History

Received 1 September 2003

Accepted after Revision 5 November 2003

Publication Date:
07 January 2004 (online)

Also available at
    Permissions and Reprints

Abstract

We developed a mouse monoclonal antibody (4G11) against insulin-like growth factor I receptor by immunizing mice with mouse embryo fibroblasts overexpressing the human insulin-like growth factor-I receptor. Not only did the 4G11 antibody inhibit the binding of [125I]insulin-like growth factor-I to the fibroblast receptor, but 4G11 antibody also potently down-regulated the insulin-like growth factor-I receptor. 4G11 Fab fragment inhibited ligand binding, but did not down-regulate the receptor, suggesting that receptor aggregation is required for down-regulation. 4G11 antibody also down-regulated the receptor in MCF-7 breast cancer cells, a panel of colon cancer cells and MG-63 osteosarcoma cells. Receptor recovery in MCF-7 cells after down-regulation by 4G11 antibody was slow, requiring 32 - 48 h for full recovery. Receptor down-regulation in MCF-7 cells by 4G11 antibody was confirmed by FACS analysis of intact and permeabilized cells. In contrast to 4G11 antibody, insulin-like growth factor-I did not down-regulate the receptor in MCF-7 cells. Down-regulation of the receptor by 4G11 antibody in MCF-7 cells resulted in inhibition of Akt and MAPK activation by insulin-like growth factor-I. We conclude that the ability of a monoclonal antibody to down-regulate the receptor may be an important antibody property in targeting the insulin-like growth factor-I receptor for the treatment of certain cancers.

Key words

Colon cancer cells - Akt - MAPK

References

  • 1 Werner H, LeRoith D. The insulin-like growth factor-I receptor signaling pathways are important for tumorigenesis and inhibition of apoptosis.  Critical Reviews in Oncogenesis. 1997;  8 71-92
    PubMedGoogle Scholar
  • 2 Baserga R, Hongo A, Rubini M, Prisco M, Valentinis B. The IGF-I receptor in cell growth, transformation and apoptosis.  Biochim Biophys Acta. 1997;  1332 F105-126
    PubMedGoogle Scholar
  • 3 Burtscher I, Christofori G. The IGF/IGF-I receptor signaling pathway as a potential target for cancer therapy.  Drug Resistance Updates. 1999;  2 3-8
    CrossrefPubMedGoogle Scholar
  • 4 Pollak M. Insulin-like growth factor physiology and cancer risk.  Eur J Cancer. 2000;  36 1224-1228
    CrossrefPubMedGoogle Scholar
  • 5 Khandwala H M, McCutcheon I E, Flyvbjerg A, Friend K E. The effects of insulin-like growth factors on tumorigenesis and neoplastic growth.  Endocrine Reviews. 2000;  21 215-244
    CrossrefPubMedGoogle Scholar
  • 6 Fisher D E. Apoptosis in cancer therapy: crossing the threshold.  Cell. 1994;  78 539-542
    CrossrefPubMedGoogle Scholar
  • 7 Turner B C, Haffty B G, Narayanan L, Yuan J, Havre P A, Gumbs A A, Kaplan L, Burgaud J-L, Carter D, Baserga R, Glazer P M. Insulin-like growth factor-I receptor overexpression mediates cellular radioresistance and local breast cancer recurrence after lumpectomy and radiation.  Cancer Res. 1997;  57 3079-3083
    PubMedGoogle Scholar
  • 8 Macaulay V M, Salisbury A J, Bohula E A, Playford M P, Smorodinsky N I, Shiloh Y. Downregulation of the type 1 insulin-like growth factor in mouse melanoma cells is associated with enhanced radiosensitivity and impaired activation of Atm kinase.  Oncogene. 2001;  20 4029-4040
    CrossrefPubMedGoogle Scholar
  • 9 Dunn S E, Hardman R A, Kari F W, Barrett J C. Insulin-like growth factor 1 (IGF-1) alters drug sensitivity of HBL100 human breast cancer cells by inhibition of apoptosis induced by diverse anticancer drugs.  Cancer Res. 1997;  57 2687-2693
    PubMedGoogle Scholar
  • 10 Lamm G M, Christofori G. Impairment of survival factor function potentiates chemotherapy-induced apoptosis in tumor cells.  Cancer Res. 1998;  58 801-807
    PubMedGoogle Scholar
  • 11 Benini S, Manara M C, Baldini N, Cerisano V, Serra M, Mercuri M, Lollini P-L, Nanni P, Picci P, Scotlandi K. Inhibition of insulin-like growth factor I receptor increases the antitumor activity of doxorubicin and vincristine against Ewing’s sarcoma cells.  Clin Cancer Res. 2001;  7 1790-1797
    PubMedGoogle Scholar
  • 12 Resnicoff M, Coppola D, Sell C, Rubin R, Ferrone S, Baserga R. Growth inhibition of human melanoma cells in nude mice by antisense strategies to the type 1 insulin-like growth factor receptor.  Cancer Res. 1994;  54 4848-4850
    PubMedGoogle Scholar
  • 13 Shapiro D N, Jones B G, Shapiro L H, Dias P, Houghton P J. Antisense-mediated reduction in insulin-like growth factor-I receptor expression suppresses the malignant phenotype of a human alveolar rhabdomyosarcoma.  J Clin Invest. 1994;  94 1235-1242
    PubMedGoogle Scholar
  • 14 Lee C-T, Wu S, Gabrilovich D, Chen H, Nadaf-Rahrov S, Ciernik I F, Carbone D P. Antitumor effects of an adenovirus expressing antisense insulin-like growth factor I receptor on human lung cancer cell lines.  Cancer Res. 1996;  56 3038-3041
    PubMedGoogle Scholar
  • 15 Chernicky D L, Yi L, Tan H, Gan S U, Ilan J. Treatment of human breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor inhibits cell growth, suppresses tumorigenesis, alters the metastatic potential, and prolongs survival in vivo.  Cancer Gene Therapy. 2000;  7 384-395
    CrossrefPubMedGoogle Scholar
  • 16 Nakamura K, Hongo A, Kodama J, Miyagi Y, Yoshinouchi M, Kudo T. Down-regulation of the insulin-like growth factor I receptor by antisense RNA can reverse the transformed phenotype of human cervical cancer cell lines.  Cancer Res. 2000;  60 760-765
    PubMedGoogle Scholar
  • 17 Scotlandi K, Avnet S, Benini S, Manara M C, Serra M, Cerisano V, Perdichizzi S, Lollini P-L, de Giovanni C, Landuzzi L, Picci P. Expression of an IGF-I receptor dominant negative mutant induces apoptosis, inhibits tumorigenesis and enhances chemosensitivity in Ewing’s sarcoma cells.  Int J Cancer. 2002;  101 11-16
    CrossrefPubMedGoogle Scholar
  • 18 Jiang Y, Rom W N, Yie T-A, Chi C X, Tchou-Wong K-M. Induction of tumor suppression and glandular differentiation of A549 lung carcinoma cells by dominant-negative IGF-I receptor.  Oncogene. 1999;  18 6071-6077
    CrossrefPubMedGoogle Scholar
  • 19 Prager D, Li H-L, Asa S, Melmed S. Dominant negative inhibition of tumorigenesis in vivo by human insulin-like growth factor I receptor mutant.  Proc Natl Acad Sci. 1994;  91 2181-2185
    PubMedGoogle Scholar
  • 20 Kalebic T, Blakesley V, Slade C, Plasschaert S, LeRoith D, Helman L J. Expression of a kinase-deficient IGF-I-R suppresses tumorigenicity of rhabdomyosarcoma cells constitutively expressing a wild type IGF-I-R.  Int J Cancer. 1998;  76 223-227
    CrossrefPubMedGoogle Scholar
  • 21 Dunn S E, Ehrlich M, Sharp N JH, Reiss K, Solomon G, Hawkins R, Baserga R, Barrett J C. A dominant negative mutant of the insulin-like growth factor-I receptor inhibits the adhesion, invasion, and metastasis of breast cancer.  Cancer Res. 1998;  58 3353-3361
    PubMedGoogle Scholar
  • 22 Adachi Y, Lee C-T, Coffee K, Yamagata N, Ohm J E, Park K-H, Dikov M M, Nadaf S R, Arteaga C L, Carbone D P. Effects of genetic blockade of the insulin-like growth factor receptor in human colon cancer cell lines.  Gastroenterology. 2002;  123 1191-1204
    CrossrefPubMedGoogle Scholar
  • 23 Arteaga C L, Kitten L J, Coronado E B, Jacobs S, Kull F C Jr, Allred D C, Osborne C K. Blockade of the type I somatomedin receptor inhibits growth of human breast cancer cells in athymic mice.  J Clin Invest. 1989;  84 1418-1423
    PubMedGoogle Scholar
  • 24 Gansler T, Furlanetto R, Gramling T S, Robinson K A, Blocker N, Buse M G, Sens D A, Garvin A J. Antibody to type I insulinlike growth factor receptor inhibits growth of Wilms’ tumor in culture and in athymic mice.  Am J Path. 1989;  135 961-966
    PubMedGoogle Scholar
  • 25 Furlanetto R W, Harwell S E, Baggs R B. Effects of insulin-like growth factor receptor inhibition on human melanomas in culture and in athymic mice.  Cancer Res. 1993;  53 2522-2526
    PubMedGoogle Scholar
  • 26 Zia F, Jacobs S, Kull F Jr, Cuttitta F, Mulshine J L, Moody T W. Monoclonal antibody αIR-3 inhibits non-small cell lung cancer growth in vitro and in vivo.  J Cellular Biochem Supp. 1996;  24 269-275
    PubMedGoogle Scholar
  • 27 Scotlandi K, Benini S, Nanni P, Lollini P-L, Nicoletti G, Landuzzi L, Serra M, Manara M C, Picci P, Baldini N. Blockage of Insulin-like growth factor-I receptor inhibits the growth of Ewing’s sarcoma in athymic mice.  Cancer Res. 1998;  58 4127-4131
    PubMedGoogle Scholar
  • 28 Kalebic T, Tsokos M, Helman L J. In vivo treatment with antibody against IGF-1 receptor suppresses growth of human rhabdomyosarcoma and down-regulates p34cdc2.  Cancer Res. 1994;  54 5531-5534
    PubMedGoogle Scholar
  • 29 Sachdev D, Li S-L, Hartell J S, Fujita-Yamaguchi Y, Miller S, Yee D. A chimeric humanized single-chain antibody against the type I insulin-like growth factor (IGF) receptor renders breast cancer cells refractory to the mitogenic effects of IGF-I.  Cancer Res. 2003;  63 627-635
    PubMedGoogle Scholar
  • 30 Hailey J, Maxwell E, Koukouras K, Bishop W R, Pachter J A, Wang Y. Neutralizing anti-insulin-like growth factor receptor 1 antibodies inhibit receptor function and induce receptor degradation in tumor cells.  Mol Cancer Therapeutics. 2002;  1 1349-1353
    PubMedGoogle Scholar
  • 31 Harlow E, Lane D (eds). Antibodies. A Laboratory Manual. Cold Spring Harbor Laboratory 1998
    Google Scholar
  • 32 Siebler T, Lopaczynski W, Terry C L, Casella S J, Munson P, De Leon D, Phang L, Blakemore K J, McEvoy R C, Kelley R I, Nissley P. Insulin-like growth factor I receptor expression and function in fibroblasts from two patients with deletion of the long arm of chromosome 15.  J Clin Endocrinol Metab. 1995;  80 3447-3457
    CrossrefPubMedGoogle Scholar
  • 33 Francis G L, Ross M, Ballard F J, Milner S J, Senn C, McNeil K A, Wallace J C, King R, Wells J RE. Novel recombinant fusion protein analogues of insulin-like growth factor (IGF)-I indicate the relative importance of IGF-binding protein and receptor binding for enhanced biological potency.  J Mol Endocrinol. 1992;  8 213-223
    PubMedGoogle Scholar
  • 34 Bostedt K T, Schmid C, Ghirlanda-Keller C, Olie R, Winterhalter K H, Zapf J. Insulin-like growth factor (IGF) I down-regulates type 1 IGF receptor (IGF 1R) and reduces the IGF I response in A549 non-small-cell lung cancer and Saos-2/B-10 osteoblastic osteosarcoma cells.  Exp Cell Res. 2001;  271 368-377
    CrossrefPubMedGoogle Scholar
  • 35 Willingham M C, Hanover J A, Dickson R B, Pastan I. Morphologic characterization of the pathway of transferrin endocytosis and recycling in human KB cells.  Proc Natl Acad Sci USA. 1984;  81 175-179
    PubMedGoogle Scholar
  • 36 Drebin J A, Link V C, Stern D F, Weinberg R A, Greene M I. Down-modulation of an oncogene protein product and reversion of the transformed phenotype by monoclonal antibodies.  Cell. 1985;  41 695-706
    CrossrefPubMedGoogle Scholar
  • 37 Maier L A, Xu F J, Hester S, Boyer C M, McKenzie S, Bruskin A M, Argon Y, Bast R C Jr. Requirements for the internalization of a murine monoclonal antibody directed against the HER-2/neu gene product c-erbB-2.  Cancer Res. 1991;  51 5361-5369
    PubMedGoogle Scholar
  • 38 Slamon D J, Leyland-Jones B, Shak S, Fuchs H, Paton V, Bajamonde A, Fleming T, Eiermann W, Wolter J, Pegram M, Baselga J, Norton L. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2.  N Engl J Med. 2001;  344 783-792
    CrossrefPubMedGoogle Scholar
  • 39 Stancovski I, Hurwitz E, Leitner O, Ullrich A, Yarden Y, Sela M. Mechanistic aspects of the opposing effects of monoclonal antibodies to the ERBB2 receptor on tumor growth.  Proc Natl Acad Sci USA. 1991;  88 8691-8695
    PubMedGoogle Scholar
  • 40 Xu F J, Boyer C M, Bae D S, Wu S, Greenwald M, O’Briant K, Yu Y H, Mills G B, Bast R C Jr. The tyrosine kinase activity of the C-erbB-2 gene product (p185) is required for growth inhibition by anti-p185 antibodies but not for the cytotoxicity of an anti-p185-Ricin-A chain immunotoxin.  Int J Cancer. 1994;  59 242-247
    PubMedGoogle Scholar
  • 41 Shawver L K, Mann E, Elliger S S, Dugger T C, Arteaga C L. Ligand-like effects induced by anti-c-erbB-2 antibodies do not correlate with and are not required for growth inhibition of human carcinoma cells.  Cancer Res. 1994;  54 1367-1373
    PubMedGoogle Scholar

P. Nissley

National Institutes of Health

Bldg 10, Rm 4N115 · Bethesda · MD 20892 · USA ·

Fax: + 1 (301) 496-9956

Email: spniss@mail.nih.gov

      >