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Logo of jcinvestThe Journal of Clinical Investigation
J Clin Invest. 1995 April; 95(4): 1897–1905.
PMCID: PMC295734

Apoptosis induced by an anti-epidermal growth factor receptor monoclonal antibody in a human colorectal carcinoma cell line and its delay by insulin.


Both EGF and insulin, or IGF, stimulate the growth of many cell types by activating receptors that contain tyrosine kinase activities. A monoclonal antibody (mAb 225) against the EGF receptor produced in this laboratory has been shown to competitively inhibit EGF binding and block activation of receptor tyrosine kinase. Here we report that a human colorectal carcinoma cell line, DiFi, which expresses high levels of EGF receptors on plasma membranes, can be induced to undergo G1 cell cycle arrest and programmed cell death (apoptosis) when cultured with mAb 225 at concentrations that saturate EGF receptors. Addition of IGF-1 or high concentrations of insulin can delay apoptosis induced by mAb 225, while the G1 arrest cannot be reversed by either IGF-1 or insulin. Insulin/IGF-1 cannot activate EGF receptor tyrosine kinase that has been inhibited by mAb 225. Moreover, an mAb against the IGF-1 receptor, which has little direct effect on DiFi cell growth, can block the capacity of insulin/IGF-1 to delay apoptosis induced by mAb 225, suggesting that the insulin/IGF-1-mediated delay of apoptosis is acting through the IGF-1 receptor. In contrast, insulin/IGF-1 cannot delay the apoptosis caused by the DNA damaging agent, cisplatin. The results indicate that EGF receptor activation is required both for cell cycle progression and for prevention of apoptosis in DiFi cells, and that a signal transduction pathway shared by receptors for insulin/IGF-1 and EGF may be involved in regulating apoptosis triggered by blockade of the EGF receptor.

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  • Carpenter G. Receptors for epidermal growth factor and other polypeptide mitogens. Annu Rev Biochem. 1987;56:881–914. [PubMed]
  • Ozanne B, Richards CS, Hendler F, Burns D, Gusterson B. Over-expression of the EGF receptor is a hallmark of squamous cell carcinomas. J Pathol. 1986 May;149(1):9–14. [PubMed]
  • Sobol RE, Astarita RW, Hofeditz C, Masui H, Fairshter R, Royston I, Mendelsohn J. Epidermal growth factor receptor expression in human lung carcinomas defined by a monoclonal antibody. J Natl Cancer Inst. 1987 Sep;79(3):403–407. [PubMed]
  • Veale D, Kerr N, Gibson GJ, Harris AL. Characterization of epidermal growth factor receptor in primary human non-small cell lung cancer. Cancer Res. 1989 Mar 1;49(5):1313–1317. [PubMed]
  • Libermann TA, Razon N, Bartal AD, Yarden Y, Schlessinger J, Soreq H. Expression of epidermal growth factor receptors in human brain tumors. Cancer Res. 1984 Feb;44(2):753–760. [PubMed]
  • Sainsbury JR, Malcolm AJ, Appleton DR, Farndon JR, Harris AL. Presence of epidermal growth factor receptor as an indicator of poor prognosis in patients with breast cancer. J Clin Pathol. 1985 Nov;38(11):1225–1228. [PMC free article] [PubMed]
  • Eisbruch A, Blick M, Lee JS, Sacks PG, Gutterman J. Analysis of the epidermal growth factor receptor gene in fresh human head and neck tumors. Cancer Res. 1987 Jul 1;47(13):3603–3605. [PubMed]
  • Neal DE, Marsh C, Bennett MK, Abel PD, Hall RR, Sainsbury JR, Harris AL. Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours. Lancet. 1985 Feb 16;1(8425):366–368. [PubMed]
  • Veale D, Ashcroft T, Marsh C, Gibson GJ, Harris AL. Epidermal growth factor receptors in non-small cell lung cancer. Br J Cancer. 1987 May;55(5):513–516. [PMC free article] [PubMed]
  • Mendelsohn J. The epidermal growth factor receptor as a target for therapy with antireceptor monoclonal antibodies. Semin Cancer Biol. 1990 Oct;1(5):339–344. [PubMed]
  • Sato JD, Kawamoto T, Le AD, Mendelsohn J, Polikoff J, Sato GH. Biological effects in vitro of monoclonal antibodies to human epidermal growth factor receptors. Mol Biol Med. 1983 Dec;1(5):511–529. [PubMed]
  • Kawamoto T, Sato JD, Le A, Polikoff J, Sato GH, Mendelsohn J. Growth stimulation of A431 cells by epidermal growth factor: identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1337–1341. [PubMed]
  • Kawamoto T, Mendelsohn J, Le A, Sato GH, Lazar CS, Gill GN. Relation of epidermal growth factor receptor concentration to growth of human epidermoid carcinoma A431 cells. J Biol Chem. 1984 Jun 25;259(12):7761–7766. [PubMed]
  • Gill GN, Kawamoto T, Cochet C, Le A, Sato JD, Masui H, McLeod C, Mendelsohn J. Monoclonal anti-epidermal growth factor receptor antibodies which are inhibitors of epidermal growth factor binding and antagonists of epidermal growth factor binding and antagonists of epidermal growth factor-stimulated tyrosine protein kinase activity. J Biol Chem. 1984 Jun 25;259(12):7755–7760. [PubMed]
  • Arteaga CL, Coronado E, Osborne CK. Blockade of the epidermal growth factor receptor inhibits transforming growth factor alpha-induced but not estrogen-induced growth of hormone-dependent human breast cancer. Mol Endocrinol. 1988 Nov;2(11):1064–1069. [PubMed]
  • Ennis BW, Valverius EM, Bates SE, Lippman ME, Bellot F, Kris R, Schlessinger J, Masui H, Goldenberg A, Mendelsohn J, et al. Anti-epidermal growth factor receptor antibodies inhibit the autocrine-stimulated growth of MDA-468 human breast cancer cells. Mol Endocrinol. 1989 Nov;3(11):1830–1838. [PubMed]
  • Karnes WE, Jr, Walsh JH, Wu SV, Kim RS, Martin MG, Wong HC, Mendelsohn J, Park JG, Cuttitta F. Autonomous proliferation of colon cancer cells that coexpress transforming growth factor alpha and its receptor. Variable effects of receptor-blocking antibody. Gastroenterology. 1992 Feb;102(2):474–485. [PubMed]
  • Reiss M, Stash EB, Vellucci VF, Zhou ZL. Activation of the autocrine transforming growth factor alpha pathway in human squamous carcinoma cells. Cancer Res. 1991 Dec 1;51(23 Pt 1):6254–6262. [PubMed]
  • Atlas I, Mendelsohn J, Baselga J, Fair WR, Masui H, Kumar R. Growth regulation of human renal carcinoma cells: role of transforming growth factor alpha. Cancer Res. 1992 Jun 15;52(12):3335–3339. [PubMed]
  • Hofer DR, Sherwood ER, Bromberg WD, Mendelsohn J, Lee C, Kozlowski JM. Autonomous growth of androgen-independent human prostatic carcinoma cells: role of transforming growth factor alpha. Cancer Res. 1991 Jun 1;51(11):2780–2785. [PubMed]
  • Mendelsohn J, Masui H, Goldenberg A. Anti-epidermal growth factor receptor monoclonal antibodies may inhibit A431 tumor cell proliferation by blocking an autocrine pathway. Trans Assoc Am Physicians. 1987;100:173–178. [PubMed]
  • Van de Vijver MJ, Kumar R, Mendelsohn J. Ligand-induced activation of A431 cell epidermal growth factor receptors occurs primarily by an autocrine pathway that acts upon receptors on the surface rather than intracellularly. J Biol Chem. 1991 Apr 25;266(12):7503–7508. [PubMed]
  • Masui H, Kawamoto T, Sato JD, Wolf B, Sato G, Mendelsohn J. Growth inhibition of human tumor cells in athymic mice by anti-epidermal growth factor receptor monoclonal antibodies. Cancer Res. 1984 Mar;44(3):1002–1007. [PubMed]
  • Masui H, Moroyama T, Mendelsohn J. Mechanism of antitumor activity in mice for anti-epidermal growth factor receptor monoclonal antibodies with different isotypes. Cancer Res. 1986 Nov;46(11):5592–5598. [PubMed]
  • Goldenberg A, Masui H, Divgi C, Kamrath H, Pentlow K, Mendelsohn J. Imaging of human tumor xenografts with an indium-111-labeled anti-epidermal growth factor receptor monoclonal antibody. J Natl Cancer Inst. 1989 Nov 1;81(21):1616–1625. [PubMed]
  • Divgi CR, Welt S, Kris M, Real FX, Yeh SD, Gralla R, Merchant B, Schweighart S, Unger M, Larson SM, et al. Phase I and imaging trial of indium 111-labeled anti-epidermal growth factor receptor monoclonal antibody 225 in patients with squamous cell lung carcinoma. J Natl Cancer Inst. 1991 Jan 16;83(2):97–104. [PubMed]
  • Rinderknecht E, Humbel RE. The amino acid sequence of human insulin-like growth factor I and its structural homology with proinsulin. J Biol Chem. 1978 Apr 25;253(8):2769–2776. [PubMed]
  • Kasuga M, Van Obberghen E, Nissley SP, Rechler MM. Demonstration of two subtypes of insulin-like growth factor receptors by affinity cross-linking. J Biol Chem. 1981 Jun 10;256(11):5305–5308. [PubMed]
  • Kasuga M, Karlsson FA, Kahn CR. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. [PubMed]
  • Jacobs S, Kull FC, Jr, Earp HS, Svoboda ME, Van Wyk JJ, Cuatrecasas P. Somatomedin-C stimulates the phosphorylation of the beta-subunit of its own receptor. J Biol Chem. 1983 Aug 25;258(16):9581–9584. [PubMed]
  • Girault JA, Chamak B, Bertuzzi G, Tixier H, Wang JK, Pang DT, Greengard P. Protein phosphotyrosine in mouse brain: developmental changes and regulation by epidermal growth factor, type I insulin-like growth factor, and insulin. J Neurochem. 1992 Feb;58(2):518–528. [PubMed]
  • Chen MC, Lee AT, Soll AH. Mitogenic response of canine fundic epithelial cells in short-term culture to transforming growth factor alpha and insulinlike growth factor I. J Clin Invest. 1991 May;87(5):1716–1723. [PMC free article] [PubMed]
  • Gross ME, Zorbas MA, Danels YJ, Garcia R, Gallick GE, Olive M, Brattain MG, Boman BM, Yeoman LC. Cellular growth response to epidermal growth factor in colon carcinoma cells with an amplified epidermal growth factor receptor derived from a familial adenomatous polyposis patient. Cancer Res. 1991 Mar 1;51(5):1452–1459. [PubMed]
  • Untawale S, Zorbas MA, Hodgson CP, Coffey RJ, Gallick GE, North SM, Wildrick DM, Olive M, Blick M, Yeoman LC, et al. Transforming growth factor-alpha production and autoinduction in a colorectal carcinoma cell line (DiFi) with an amplified epidermal growth factor receptor gene. Cancer Res. 1993 Apr 1;53(7):1630–1636. [PubMed]
  • Kyprianou N, English HF, Isaacs JT. Activation of a Ca2+-Mg2+-dependent endonuclease as an early event in castration-induced prostatic cell death. Prostate. 1988;13(2):103–117. [PubMed]
  • Kyprianou N, Isaacs JT. Activation of programmed cell death in the rat ventral prostate after castration. Endocrinology. 1988 Feb;122(2):552–562. [PubMed]
  • Kyprianou N, English HF, Isaacs JT. Programmed cell death during regression of PC-82 human prostate cancer following androgen ablation. Cancer Res. 1990 Jun 15;50(12):3748–3753. [PubMed]
  • Kyprianou N, Isaacs JT. Expression of transforming growth factor-beta in the rat ventral prostate during castration-induced programmed cell death. Mol Endocrinol. 1989 Oct;3(10):1515–1522. [PubMed]
  • Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972 Aug;26(4):239–257. [PMC free article] [PubMed]
  • Wyllie AH, Kerr JF, Currie AR. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. [PubMed]
  • Kolber MA, Broschat KO, Landa-Gonzalez B. Cytochalasin B induces cellular DNA fragmentation. FASEB J. 1990 Sep;4(12):3021–3027. [PubMed]
  • Waterfield MD, Mayes EL, Stroobant P, Bennet PL, Young S, Goodfellow PN, Banting GS, Ozanne B. A monoclonal antibody to the human epidermal growth factor receptor. J Cell Biochem. 1982;20(2):149–161. [PubMed]
  • Ludlow JW, DeCaprio JA, Huang CM, Lee WH, Paucha E, Livingston DM. SV40 large T antigen binds preferentially to an underphosphorylated member of the retinoblastoma susceptibility gene product family. Cell. 1989 Jan 13;56(1):57–65. [PubMed]
  • DeCaprio JA, Ludlow JW, Lynch D, Furukawa Y, Griffin J, Piwnica-Worms H, Huang CM, Livingston DM. The product of the retinoblastoma susceptibility gene has properties of a cell cycle regulatory element. Cell. 1989 Sep 22;58(6):1085–1095. [PubMed]
  • Rohlik QT, Adams D, Kull FC, Jr, Jacobs S. An antibody to the receptor for insulin-like growth factor I inhibits the growth of MCF-7 cells in tissue culture. Biochem Biophys Res Commun. 1987 Nov 30;149(1):276–281. [PubMed]
  • Sen S, D'Incalci M. Apoptosis. Biochemical events and relevance to cancer chemotherapy. FEBS Lett. 1992 Jul 27;307(1):122–127. [PubMed]
  • Andrews PA, Howell SB. Cellular pharmacology of cisplatin: perspectives on mechanisms of acquired resistance. Cancer Cells. 1990 Feb;2(2):35–43. [PubMed]
  • Stampfer MR, Pan CH, Hosoda J, Bartholomew J, Mendelsohn J, Yaswen P. Blockage of EGF receptor signal transduction causes reversible arrest of normal and immortal human mammary epithelial cells with synchronous reentry into the cell cycle. Exp Cell Res. 1993 Sep;208(1):175–188. [PubMed]
  • Markowitz SD, Molkentin K, Gerbic C, Jackson J, Stellato T, Willson JK. Growth stimulation by coexpression of transforming growth factor-alpha and epidermal growth factor-receptor in normal and adenomatous human colon epithelium. J Clin Invest. 1990 Jul;86(1):356–362. [PMC free article] [PubMed]
  • Gill GN, Lazar CS. Increased phosphotyrosine content and inhibition of proliferation in EGF-treated A431 cells. Nature. 1981 Sep 24;293(5830):305–307. [PubMed]
  • Kaplan O, Jaroszewski JW, Faustino PJ, Zugmaier G, Ennis BW, Lippman M, Cohen JS. Toxicity and effects of epidermal growth factor on glucose metabolism of MDA-468 human breast cancer cells. J Biol Chem. 1990 Aug 15;265(23):13641–13649. [PubMed]
  • Fan Z, Mendelsohn J, Masui H, Kumar R. Regulation of epidermal growth factor receptor in NIH3T3/HER14 cells by antireceptor monoclonal antibodies. J Biol Chem. 1993 Oct 5;268(28):21073–21079. [PubMed]
  • Fan Z, Lu Y, Wu X, Mendelsohn J. Antibody-induced epidermal growth factor receptor dimerization mediates inhibition of autocrine proliferation of A431 squamous carcinoma cells. J Biol Chem. 1994 Nov 4;269(44):27595–27602. [PubMed]
  • Pietrzkowski Z, Sell C, Lammers R, Ullrich A, Baserga R. Roles of insulinlike growth factor 1 (IGF-1) and the IGF-1 receptor in epidermal growth factor-stimulated growth of 3T3 cells. Mol Cell Biol. 1992 Sep;12(9):3883–3889. [PMC free article] [PubMed]

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