Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Jul;78(7):4339–4343. doi: 10.1073/pnas.78.7.4339

Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells.

R F Santerre, R A Cook, R M Crisel, J D Sharp, R J Schmidt, D C Williams, C P Wilson
PMCID: PMC319785  PMID: 6270673

Abstract

A clonal hamster beta cell line (HIT) was established by simian virus 40 transformation of Syrian hamster pancreatic islet cells. Cytoplasmic insulin was detected in all cells by indirect fluorescent antibody staining, and membrane-bound secretory granules were observed ultrastructurally. Acidified-ethanol extracts of HIT cell cultures contained hamster insulin as determined by radioimmunoassay, radioreceptor assay, and bioassay. One subclone at passage 39 contained 2.6 micrograms of insulin per mg of cell protein. [3H]Leucine-labeled HIT insulin and proinsulin were identical to islet-derived proteins when compared by NaDodSO4/polyacrylamide gel electrophoresis of immunoprecipitates. HIT cell insulin secretion was stimulated by glucose, glucagon, and 3-isobutyl-1-methylxanthine. Insulin secretion at optimal glucose concentration (7.5 mM) was 2.4 milliunits per 10(6) cells per hr. Somatostatin and dexamethasone markedly inhibited HIT insulin secretion. The HIT cell line represents a unique in vitro system for studying beta cell metabolism and insulin biosynthesis.

Full text

PDF
4341

Images in this article

4340Image
on p.4340
4341Image
on p.4341
4341Image
on p.4341
4341Image
on p.4341
4341Image
on p.4341
4341Image
on p.4341

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amherdt M., Orci L., Track N. S., Lambert A. E., Kanazawa Y., Stauffacher W. An ultrastructural study of the islet cell tumor of the golden hamster. Horm Metab Res. 1971 Jul;3(4):252–258. doi: 10.1055/s-0028-1094143. [DOI] [PubMed] [Google Scholar]
  2. Braaten J. T., Lee M. J., Schenk A., Mintz D. H. Removal of fibroblastoid cells from primary monolayer cultures of rat neonatal endocrine pancreas by sodium ethylmercurithiosalicylate. Biochem Biophys Res Commun. 1974 Nov 27;61(2):476–482. doi: 10.1016/0006-291x(74)90981-4. [DOI] [PubMed] [Google Scholar]
  3. Chick W. L. Beta cell replication in rat pancreatic monolayer cultures. Effects of glucose, tolbutamide, glucocorticoid, growth hormone and glucagon. Diabetes. 1973 Sep;22(9):687–693. doi: 10.2337/diab.22.9.687. [DOI] [PubMed] [Google Scholar]
  4. Chick W. L., Warren S., Chute R. N., Like A. A., Lauris V., Kitchen K. C. A transplantable insulinoma in the rat. Proc Natl Acad Sci U S A. 1977 Feb;74(2):628–632. doi: 10.1073/pnas.74.2.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chou J. Y. Establishment of clonal human placental cells synthesizing human choriogonadotropin. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1854–1858. doi: 10.1073/pnas.75.4.1854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crook R. B., Louie M., Deuel T. F., Tomkins G. M. Regulation of glutamine synthetase by dexamethasone in hepatoma tissue culture cells. J Biol Chem. 1978 Sep 10;253(17):6125–6131. [PubMed] [Google Scholar]
  7. Duguid J. R., Steiner D. F., Chick W. L. Partial purification and characterization of the mRNA for rat preproinsulin. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3539–3543. doi: 10.1073/pnas.73.10.3539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gazdar A. F., Chick W. L., Oie H. K., Sims H. L., King D. L., Weir G. C., Lauris V. Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3519–3523. doi: 10.1073/pnas.77.6.3519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ide T., Anzai K., Andoh T. Enhancement of SV40 transformation by treatment of C3H2K cells with UV light and caffeine. I. Combined effect of UV light and caffeine. Virology. 1975 Aug;66(2):568–578. doi: 10.1016/0042-6822(75)90229-9. [DOI] [PubMed] [Google Scholar]
  10. KIRKMAN H. A preliminary report concerning tumors observed in Syrian hamsters. Stanford Med Bull. 1962 Nov;20:163–166. [PubMed] [Google Scholar]
  11. Kostianovsky M., McDaniel M. L., Still M. F., Codilla R. C., Lacy P. E. Monolayer cell culture of adult rat islets of Langerhans. Diabetologia. 1974 Aug;10(4):337–344. doi: 10.1007/BF02627736. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lacy P. E., Kostianovsky M. Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes. 1967 Jan;16(1):35–39. doi: 10.2337/diab.16.1.35. [DOI] [PubMed] [Google Scholar]
  14. Lambert A. E., Blondel B., Kanazawa Y., Orci L., Renold A. E. Monolayer cell culture of neonatal rat pancreas: light microscopy and evidence for immunoreactive insulin synthesis and release. Endocrinology. 1972 Jan;90(1):239–248. doi: 10.1210/endo-90-1-239. [DOI] [PubMed] [Google Scholar]
  15. Laskey R. A., Mills A. D. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film. FEBS Lett. 1977 Oct 15;82(2):314–316. doi: 10.1016/0014-5793(77)80609-1. [DOI] [PubMed] [Google Scholar]
  16. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  17. Lomedico P. T., Saunders G. F. Preparation of pancreatic mRNA: cell-free translation of an insulin-immunoreactive polypeptide. Nucleic Acids Res. 1976 Feb;3(2):381–391. doi: 10.1093/nar/3.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marliss E. B., Wollheim C. B., Blondel B., Orci L., Lambert A. E., Stauffacher W., Like A. A., Renold A. E. Insulin and glucagon release from monolayer cell cultures of pancreas from newborn rats. Eur J Clin Invest. 1973 Jan;3(1):16–26. doi: 10.1111/j.1365-2362.1973.tb00324.x. [DOI] [PubMed] [Google Scholar]
  19. Niesor E. J., Wollheim C. B., Mintz D. H., Blondel B., Renold A. E., Weil R. Establishment of rat pancreatic endocrine cell lines by infection with simian virus 40. Biochem J. 1979 Mar 15;178(3):559–568. doi: 10.1042/bj1780559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. POPE J. H., ROWE W. P. DETECTION OF SPECIFIC ANTIGEN IN SV40-TRANSFORMED CELLS BY IMMUNOFLUORESCENCE. J Exp Med. 1964 Aug 1;120:121–128. doi: 10.1084/jem.120.2.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rae P. A., Yip C. C., Schimmer B. P. Isolation of cloned Syrian hamster insulinoma cell lines with limited capacity for insulin production. Can J Physiol Pharmacol. 1979 Aug;57(8):819–824. doi: 10.1139/y79-125. [DOI] [PubMed] [Google Scholar]
  22. Siegel E. G., Wollheim C. B., Sharp G. W., Herberg L., Renold A. E. Defective calcium handling and insulin release in islets from diabetic Chinese hamsters. Biochem J. 1979 Apr 15;180(1):233–236. doi: 10.1042/bj1800233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Simar L. J., Sodoyez J. C., Grillo A. I., Nagano T., Lefebvre P. J., Foa P. P. Biological properties of a transplantable islet-cell tumor of the golden hamster. 3. Electron microscopic studies. Diabetes. 1968 Sep;17(9):582–587. doi: 10.2337/diab.17.9.582. [DOI] [PubMed] [Google Scholar]
  24. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  25. Turcot-Lemay L., Lemay A., Lacy P. E. Somatostatin inhibition of insulin release from freshly isolated and organ cultured rat islets of Langerhans in vitro. Biochem Biophys Res Commun. 1975 Apr 21;63(4):1130–1138. doi: 10.1016/0006-291x(75)90686-5. [DOI] [PubMed] [Google Scholar]
  26. Uchida S., Watanabe S., Aizawa T., Furuno A., Muto T. Polyoncogenicity and insulinoma-inducing ability of BK Virus, a human Papovavirus, in Syrian golden hamsters. J Natl Cancer Inst. 1979 Jul;63(1):119–126. [PubMed] [Google Scholar]
  27. Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wacker A., Kaul S., Eichler A. R. Insulin-Synthese fusionierter tierischer Zellen in Suspensionkultur. Naturwissenschaften. 1978 Jan;65(1):69–70. doi: 10.1007/BF00420649. [DOI] [PubMed] [Google Scholar]
  29. Watanabe H., Nicholson W. E., Orth D. N. Inhibition of adrenocorticotropic hormone production by glucocorticoids in mouse pituitary tumor cells. Endocrinology. 1973 Aug;93(2):411–416. doi: 10.1210/endo-93-2-411. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES