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. 1989 Sep;84(3):829–839. doi: 10.1172/JCI114243

Autonomous growth of a human neuroblastoma cell line is mediated by insulin-like growth factor II.

O M El-Badry 1, J A Romanus 1, L J Helman 1, M J Cooper 1, M M Rechler 1, M A Israel 1
PMCID: PMC329726  PMID: 2547840

Abstract

Insulin-like growth factor II (IGF-II) mRNA was increased in two of eight neuroblastomas and in eight of eight pheochromocytomas, tumors of the adrenal medulla that occur in childhood and adulthood, respectively. RNA encoding the type I IGF receptor, the receptor thought to mediate the mitogenic effects of IGF-I and IGF-II, also was uniformly expressed in these cells. To assess the role of IGF-II in the growth of these tumor cells, we have used the SK-N-AS cultured neuroblastoma cell line, which can be continuously propagated in mitogen-free medium, as a model system. Our results strongly suggest that IGF-II, synthesized by SK-N-AS cells and acting through type I IGF receptors, contributes to the autonomous growth of this tumor cell line. (a) SK-N-AS cells synthesized large amounts of IGF-II RNA and secreted greater than 50 ng/ml of IGF-II (as determined by specific radioimmuno- and radioreceptor assays). Little, if any, IGF-I RNA or immunoreactive IGF-I were detected. (b) SK-N-AS cells possess type I IGF receptors. (c) Exogenous IGF-I and IGF-II stimulated DNA synthesis in SK-N-AS cells, and this stimulation was abolished by a blocking antibody to the type I IGF receptor. (d) This anti-receptor antibody also abolished the multiplication of SK-N-AS cells in the absence of added mitogens. We conclude that IGF-II is an autocrine growth factor for SK-N-AS cells and suggest that this mechanism may contribute to the growth of some adrenal medullary tumors.

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Selected References

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  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell G. I., Merryweather J. P., Sanchez-Pescador R., Stempien M. M., Priestley L., Scott J., Rall L. B. Sequence of a cDNA clone encoding human preproinsulin-like growth factor II. 1984 Aug 30-Sep 5Nature. 310(5980):775–777. doi: 10.1038/310775a0. [DOI] [PubMed] [Google Scholar]
  3. Brown A. L., Graham D. E., Nissley S. P., Hill D. J., Strain A. J., Rechler M. M. Developmental regulation of insulin-like growth factor II mRNA in different rat tissues. J Biol Chem. 1986 Oct 5;261(28):13144–13150. [PubMed] [Google Scholar]
  4. Cariani E., Lasserre C., Seurin D., Hamelin B., Kemeny F., Franco D., Czech M. P., Ullrich A., Brechot C. Differential expression of insulin-like growth factor II mRNA in human primary liver cancers, benign liver tumors, and liver cirrhosis. Cancer Res. 1988 Dec 1;48(23):6844–6849. [PubMed] [Google Scholar]
  5. Casella S. J., Han V. K., D'Ercole A. J., Svoboda M. E., Van Wyk J. J. Insulin-like growth factor II binding to the type I somatomedin receptor. Evidence for two high affinity binding sites. J Biol Chem. 1986 Jul 15;261(20):9268–9273. [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Clemmons D. R., Van Wyk J. J. Evidence for a functional role of endogenously produced somatomedinlike peptides in the regulation of DNA synthesis in cultured human fibroblasts and porcine smooth muscle cells. J Clin Invest. 1985 Jun;75(6):1914–1918. doi: 10.1172/JCI111906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clemmons D. R., Van Wyk J. J. Factors controlling blood concentration of somatomedin C. Clin Endocrinol Metab. 1984 Mar;13(1):113–143. doi: 10.1016/s0300-595x(84)80011-0. [DOI] [PubMed] [Google Scholar]
  9. Conover C. A., Rosenfeld R. G., Hintz R. L. Insulin-like growth factor II binding and action in human fetal fibroblasts. J Cell Physiol. 1987 Dec;133(3):560–566. doi: 10.1002/jcp.1041330318. [DOI] [PubMed] [Google Scholar]
  10. Daughaday W. H., Emanuele M. A., Brooks M. H., Barbato A. L., Kapadia M., Rotwein P. Synthesis and secretion of insulin-like growth factor II by a leiomyosarcoma with associated hypoglycemia. N Engl J Med. 1988 Dec 1;319(22):1434–1440. doi: 10.1056/NEJM198812013192202. [DOI] [PubMed] [Google Scholar]
  11. Daughaday W. H., Hall K., Salmon W. D., Jr, Van den Brande J. L., Van Wyk J. J. On the nomenclature of the somatomedins and insulin-like growth factors. Endocrinology. 1987 Nov;121(5):1911–1912. doi: 10.1210/endo-121-5-1911. [DOI] [PubMed] [Google Scholar]
  12. Denizot F., Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods. 1986 May 22;89(2):271–277. doi: 10.1016/0022-1759(86)90368-6. [DOI] [PubMed] [Google Scholar]
  13. Ewton D. Z., Falen S. L., Florini J. R. The type II insulin-like growth factor (IGF) receptor has low affinity for IGF-I analogs: pleiotypic actions of IGFs on myoblasts are apparently mediated by the type I receptor. Endocrinology. 1987 Jan;120(1):115–123. doi: 10.1210/endo-120-1-115. [DOI] [PubMed] [Google Scholar]
  14. Flier J. S., Usher P., Moses A. C. Monoclonal antibody to the type I insulin-like growth factor (IGF-I) receptor blocks IGF-I receptor-mediated DNA synthesis: clarification of the mitogenic mechanisms of IGF-I and insulin in human skin fibroblasts. Proc Natl Acad Sci U S A. 1986 Feb;83(3):664–668. doi: 10.1073/pnas.83.3.664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fu X. X., Su C. Y., Lee Y., Hintz R., Biempica L., Snyder R., Rogler C. E. Insulinlike growth factor II expression and oval cell proliferation associated with hepatocarcinogenesis in woodchuck hepatitis virus carriers. J Virol. 1988 Sep;62(9):3422–3430. doi: 10.1128/jvi.62.9.3422-3430.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Furlanetto R. W., DiCarlo J. N., Wisehart C. The type II insulin-like growth factor receptor does not mediate deoxyribonucleic acid synthesis in human fibroblasts. J Clin Endocrinol Metab. 1987 Jun;64(6):1142–1149. doi: 10.1210/jcem-64-6-1142. [DOI] [PubMed] [Google Scholar]
  17. Furlanetto R. W., Underwood L. E., Van Wyk J. J., D'Ercole A. J. Estimation of somatomedin-C levels in normals and patients with pituitary disease by radioimmunoassay. J Clin Invest. 1977 Sep;60(3):648–657. doi: 10.1172/JCI108816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gowan L. K., Hampton B., Hill D. J., Schlueter R. J., Perdue J. F. Purification and characterization of a unique high molecular weight form of insulin-like growth factor II. Endocrinology. 1987 Aug;121(2):449–458. doi: 10.1210/endo-121-2-449. [DOI] [PubMed] [Google Scholar]
  19. Gray A., Tam A. W., Dull T. J., Hayflick J., Pintar J., Cavenee W. K., Koufos A., Ullrich A. Tissue-specific and developmentally regulated transcription of the insulin-like growth factor 2 gene. DNA. 1987 Aug;6(4):283–295. doi: 10.1089/dna.1987.6.283. [DOI] [PubMed] [Google Scholar]
  20. Greenstein L. A., Gaynes L. A., Romanus J. A., Lee L., Rechler M. M., Nissley S. P. Purification of rat insulin-like growth factor II. Methods Enzymol. 1987;146:259–269. doi: 10.1016/s0076-6879(87)46028-x. [DOI] [PubMed] [Google Scholar]
  21. Han V. K., D'Ercole A. J., Lund P. K. Cellular localization of somatomedin (insulin-like growth factor) messenger RNA in the human fetus. Science. 1987 Apr 10;236(4798):193–197. doi: 10.1126/science.3563497. [DOI] [PubMed] [Google Scholar]
  22. Han V. K., Lund P. K., Lee D. C., D'Ercole A. J. Expression of somatomedin/insulin-like growth factor messenger ribonucleic acids in the human fetus: identification, characterization, and tissue distribution. J Clin Endocrinol Metab. 1988 Feb;66(2):422–429. doi: 10.1210/jcem-66-2-422. [DOI] [PubMed] [Google Scholar]
  23. Haselbacher G. K., Irminger J. C., Zapf J., Ziegler W. H., Humbel R. E. Insulin-like growth factor II in human adrenal pheochromocytomas and Wilms tumors: expression at the mRNA and protein level. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1104–1106. doi: 10.1073/pnas.84.4.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Helman L. J., Thiele C. J., Linehan W. M., Nelkin B. D., Baylin S. B., Israel M. A. Molecular markers of neuroendocrine development and evidence of environmental regulation. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2336–2339. doi: 10.1073/pnas.84.8.2336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Helson L., Member B., Helson C. Importance of clinical exposure on verapamil enhancement of adriamycin-vincristine cytotoxicity in human neuroblastoma. Cancer Drug Deliv. 1984 Fall;1(4):303–305. doi: 10.1089/cdd.1984.1.303. [DOI] [PubMed] [Google Scholar]
  26. Huff K. K., Kaufman D., Gabbay K. H., Spencer E. M., Lippman M. E., Dickson R. B. Secretion of an insulin-like growth factor-I-related protein by human breast cancer cells. Cancer Res. 1986 Sep;46(9):4613–4619. [PubMed] [Google Scholar]
  27. Höppener J. W., Mosselman S., Roholl P. J., Lambrechts C., Slebos R. J., de Pagter-Holthuizen P., Lips C. J., Jansz H. S., Sussenbach J. S. Expression of insulin-like growth factor-I and -II genes in human smooth muscle tumours. EMBO J. 1988 May;7(5):1379–1385. doi: 10.1002/j.1460-2075.1988.tb02954.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Höppener J. W., Steenbergh P. H., Slebos R. J., de Pagter-Holthuizen P., Roos B. A., Jansen M., Van den Brande J. L., Sussenbach J. S., Jansz H. S., Lips C. J. Expression of insulin-like growth factor-I and -II genes in rat medullary thyroid carcinoma. FEBS Lett. 1987 May 4;215(1):122–126. doi: 10.1016/0014-5793(87)80125-4. [DOI] [PubMed] [Google Scholar]
  29. Irminger J. C., Rosen K. M., Humbel R. E., Villa-Komaroff L. Tissue-specific expression of insulin-like growth factor II mRNAs with distinct 5' untranslated regions. Proc Natl Acad Sci U S A. 1987 Sep;84(18):6330–6334. doi: 10.1073/pnas.84.18.6330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Israel M. A. The evolution of clinical molecular genetics. Neuroblastoma as a model tumor. Am J Pediatr Hematol Oncol. 1986 Summer;8(2):163–172. [PubMed] [Google Scholar]
  31. Jansen M., van Schaik F. M., Ricker A. T., Bullock B., Woods D. E., Gabbay K. H., Nussbaum A. L., Sussenbach J. S., Van den Brande J. L. Sequence of cDNA encoding human insulin-like growth factor I precursor. Nature. 1983 Dec 8;306(5943):609–611. doi: 10.1038/306609a0. [DOI] [PubMed] [Google Scholar]
  32. Keating M. T., Williams L. T. Autocrine stimulation of intracellular PDGF receptors in v-sis-transformed cells. Science. 1988 Feb 19;239(4842):914–916. doi: 10.1126/science.2829358. [DOI] [PubMed] [Google Scholar]
  33. Kiess W., Blickenstaff G. D., Sklar M. M., Thomas C. L., Nissley S. P., Sahagian G. G. Biochemical evidence that the type II insulin-like growth factor receptor is identical to the cation-independent mannose 6-phosphate receptor. J Biol Chem. 1988 Jul 5;263(19):9339–9344. [PubMed] [Google Scholar]
  34. Kiess W., Haskell J. F., Lee L., Greenstein L. A., Miller B. E., Aarons A. L., Rechler M. M., Nissley S. P. An antibody that blocks insulin-like growth factor (IGF) binding to the type II IGF receptor is neither an agonist nor an inhibitor of IGF-stimulated biologic responses in L6 myoblasts. J Biol Chem. 1987 Sep 15;262(26):12745–12751. [PubMed] [Google Scholar]
  35. Kiess W., Thomas C. L., Greenstein L. A., Lee L., Sklar M. M., Rechler M. M., Sahagian G. G., Nissley S. P. Insulin-like growth factor-II (IGF-II) inhibits both the cellular uptake of beta-galactosidase and the binding of beta-galactosidase to purified IGF-II/mannose 6-phosphate receptor. J Biol Chem. 1989 Mar 15;264(8):4710–4714. [PubMed] [Google Scholar]
  36. Kull F. C., Jr, Jacobs S., Su Y. F., Svoboda M. E., Van Wyk J. J., Cuatrecasas P. Monoclonal antibodies to receptors for insulin and somatomedin-C. J Biol Chem. 1983 May 25;258(10):6561–6566. [PubMed] [Google Scholar]
  37. Maciel R. M., Moses A. C., Villone G., Tramontano D., Ingbar S. H. Demonstration of the production and physiological role of insulin-like growth factor II in rat thyroid follicular cells in culture. J Clin Invest. 1988 Nov;82(5):1546–1553. doi: 10.1172/JCI113764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Morgan D. O., Edman J. C., Standring D. N., Fried V. A., Smith M. C., Roth R. A., Rutter W. J. Insulin-like growth factor II receptor as a multifunctional binding protein. Nature. 1987 Sep 24;329(6137):301–307. doi: 10.1038/329301a0. [DOI] [PubMed] [Google Scholar]
  39. Moses A. C., Nissley S. P., Short P. A., Rechler M. M., White R. M., Knight A. B., Higa O. Z. Increased levels of multiplication-stimulating activity, an insulin-like growth factor, in fetal rat serum. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3649–3653. doi: 10.1073/pnas.77.6.3649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nakanishi Y., Cuttitta F., Kasprzyk P. G., Avis I., Steinberg S. M., Gazdar A. F., Mulshine J. L. Growth factor effects on small cell lung cancer cells using a colorimetric assay: can a transferrin-like factor mediate autocrine growth? Exp Cell Biol. 1988;56(1-2):74–85. doi: 10.1159/000163465. [DOI] [PubMed] [Google Scholar]
  41. Nakanishi Y., Mulshine J. L., Kasprzyk P. G., Natale R. B., Maneckjee R., Avis I., Treston A. M., Gazdar A. F., Minna J. D., Cuttitta F. Insulin-like growth factor-I can mediate autocrine proliferation of human small cell lung cancer cell lines in vitro. J Clin Invest. 1988 Jul;82(1):354–359. doi: 10.1172/JCI113594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Norstedt G., Levinovitz A., Möller C., Eriksson L. C., Andersson G. Expression of insulin-like growth factor I (IGF-I) and IGF-II mRNA during hepatic development, proliferation and carcinogenesis in the rat. Carcinogenesis. 1988 Feb;9(2):209–213. doi: 10.1093/carcin/9.2.209. [DOI] [PubMed] [Google Scholar]
  43. Rechler M. M., Nissley S. P. The nature and regulation of the receptors for insulin-like growth factors. Annu Rev Physiol. 1985;47:425–442. doi: 10.1146/annurev.ph.47.030185.002233. [DOI] [PubMed] [Google Scholar]
  44. Reeve A. E., Eccles M. R., Wilkins R. J., Bell G. I., Millow L. J. Expression of insulin-like growth factor-II transcripts in Wilms' tumour. Nature. 1985 Sep 19;317(6034):258–260. doi: 10.1038/317258a0. [DOI] [PubMed] [Google Scholar]
  45. Rohlik Q. T., Adams D., Kull F. C., 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. doi: 10.1016/0006-291x(87)91635-4. [DOI] [PubMed] [Google Scholar]
  46. Romanus J. A., Rabinovitch A., Rechler M. M. Neonatal rat islet cell cultures synthesize insulin-like growth factor I. Diabetes. 1985 Jul;34(7):696–702. doi: 10.2337/diab.34.7.696. [DOI] [PubMed] [Google Scholar]
  47. Roth R. A. Structure of the receptor for insulin-like growth factor II: the puzzle amplified. Science. 1988 Mar 11;239(4845):1269–1271. doi: 10.1126/science.2964085. [DOI] [PubMed] [Google Scholar]
  48. Schoenle E., Zapf J., Humbel R. E., Froesch E. R. Insulin-like growth factor I stimulates growth in hypophysectomized rats. Nature. 1982 Mar 18;296(5854):252–253. doi: 10.1038/296252a0. [DOI] [PubMed] [Google Scholar]
  49. Scott J., Cowell J., Robertson M. E., Priestley L. M., Wadey R., Hopkins B., Pritchard J., Bell G. I., Rall L. B., Graham C. F. Insulin-like growth factor-II gene expression in Wilms' tumour and embryonic tissues. Nature. 1985 Sep 19;317(6034):260–262. doi: 10.1038/317260a0. [DOI] [PubMed] [Google Scholar]
  50. Soares M. B., Turken A., Ishii D., Mills L., Episkopou V., Cotter S., Zeitlin S., Efstratiadis A. Rat insulin-like growth factor II gene. A single gene with two promoters expressing a multitranscript family. J Mol Biol. 1986 Dec 20;192(4):737–752. doi: 10.1016/0022-2836(86)90025-2. [DOI] [PubMed] [Google Scholar]
  51. Sugimoto T., Sawada T., Arakawa S., Matsumura T., Sakamoto I., Takeuchi Y., Reynolds C. P., Kemshead J. T., Helson L. Possible differential diagnosis of neuroblastoma from rhabdomyosarcoma and Ewing's sarcoma by using a panel of monoclonal antibodies. Jpn J Cancer Res. 1985 Apr;76(4):301–307. [PubMed] [Google Scholar]
  52. Sugimoto T., Tatsumi E., Kemshead J. T., Helson L., Green A. A., Minowada J. Determination of cell surface membrane antigens common to both human neuroblastoma and leukemia-lymphoma cell lines by a panel of 38 monoclonal antibodies. J Natl Cancer Inst. 1984 Jul;73(1):51–57. [PubMed] [Google Scholar]
  53. Thiele C. J., Reynolds C. P., Israel M. A. Decreased expression of N-myc precedes retinoic acid-induced morphological differentiation of human neuroblastoma. 1985 Jan 31-Feb 6Nature. 313(6001):404–406. doi: 10.1038/313404a0. [DOI] [PubMed] [Google Scholar]
  54. Tricoli J. V., Rall L. B., Karakousis C. P., Herrera L., Petrelli N. J., Bell G. I., Shows T. B. Enhanced levels of insulin-like growth factor messenger RNA in human colon carcinomas and liposarcomas. Cancer Res. 1986 Dec;46(12 Pt 1):6169–6173. [PubMed] [Google Scholar]
  55. Ueno T., Takahashi K., Matsuguchi T., Ikejiri K., Endo H., Yamamoto M. Reactivation of rat insulin-like growth factor II gene during hepatocarcinogenesis. Carcinogenesis. 1988 Oct;9(10):1779–1783. doi: 10.1093/carcin/9.10.1779. [DOI] [PubMed] [Google Scholar]
  56. Ullrich A., Gray A., Tam A. W., Yang-Feng T., Tsubokawa M., Collins C., Henzel W., Le Bon T., Kathuria S., Chen E. Insulin-like growth factor I receptor primary structure: comparison with insulin receptor suggests structural determinants that define functional specificity. EMBO J. 1986 Oct;5(10):2503–2512. doi: 10.1002/j.1460-2075.1986.tb04528.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Van Obberghen-Schilling E. E., Rechler M. M., Romanus J. A., Knight A. B., Nissley S. P., Humbel R. E. Receptors for insulinlike growth factor I are defective in fibroblasts cultured from a patient with leprechaunism. J Clin Invest. 1981 Nov;68(5):1356–1365. doi: 10.1172/JCI110383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Van Wyk J. J., Graves D. C., Casella S. J., Jacobs S. Evidence from monoclonal antibody studies that insulin stimulates deoxyribonucleic acid synthesis through the type I somatomedin receptor. J Clin Endocrinol Metab. 1985 Oct;61(4):639–643. doi: 10.1210/jcem-61-4-639. [DOI] [PubMed] [Google Scholar]
  59. Whitfield H. J., Bruni C. B., Frunzio R., Terrell J. E., Nissley S. P., Rechler M. M. Isolation of a cDNA clone encoding rat insulin-like growth factor-II precursor. Nature. 1984 Nov 15;312(5991):277–280. doi: 10.1038/312277a0. [DOI] [PubMed] [Google Scholar]
  60. Yee D., Cullen K. J., Paik S., Perdue J. F., Hampton B., Schwartz A., Lippman M. E., Rosen N. Insulin-like growth factor II mRNA expression in human breast cancer. Cancer Res. 1988 Dec 1;48(23):6691–6696. [PubMed] [Google Scholar]
  61. Yee D., Paik S., Lebovic G. S., Marcus R. R., Favoni R. E., Cullen K. J., Lippman M. E., Rosen N. Analysis of insulin-like growth factor I gene expression in malignancy: evidence for a paracrine role in human breast cancer. Mol Endocrinol. 1989 Mar;3(3):509–517. doi: 10.1210/mend-3-3-509. [DOI] [PubMed] [Google Scholar]
  62. de Pagter-Holthuizen P., Jansen M., van Schaik F. M., van der Kammen R., Oosterwijk C., Van den Brande J. L., Sussenbach J. S. The human insulin-like growth factor II gene contains two development-specific promoters. FEBS Lett. 1987 Apr 20;214(2):259–264. doi: 10.1016/0014-5793(87)80066-2. [DOI] [PubMed] [Google Scholar]
  63. de Pagter-Holthuizen P., Jansen M., van der Kammen R. A., van Schaik F. M., Sussenbach J. S. Differential expression of the human insulin-like growth factor II gene. Characterization of the IGF-II mRNAs and an mRNA encoding a putative IGF-II-associated protein. Biochim Biophys Acta. 1988 Sep 7;950(3):282–295. doi: 10.1016/0167-4781(88)90124-8. [DOI] [PubMed] [Google Scholar]

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