Skip to main content
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1996 May;87(5):451–458. doi: 10.1111/j.1349-7006.1996.tb00245.x

Establishment of a Clonal Cell Line Producing Granulocyte Colony‐Stimulating Factor and Parathyroid Hormone‐Related Protein from a Lung Cancer Patient with Leukocytosis and Hypercalcemia

Yasutomo Asahi 1, Ichiro Kubonishi 1,, Jun Imamura 1, Mikio Kamioka 1, Hiroshi Matsushita 3, Mutsuo Furihata 2, Yuji Ohtsuki 2, Isao Miyoshi 1
PMCID: PMC5921125  PMID: 8641981

Abstract

Squamous cell lung carcinoma cells obtained from a patient who presented with leukocytosis and hypercalcemia were transplanted into nude mice anda serially transplantable cell line, OKa‐N‐1, was established. The nude mice transplanted with OKa‐N‐1 cells displayed leukocytosis and hypercalcemia. Serum levels of granulocyte colony‐stimulating factor (G‐CSF) and parathyroid hormone‐related protein (PTHrP) were both elevated in these mice. In vitro cultivation of this tumor cell line gave rise to a clonal cell line, OKa‐C‐1. Nude mice transplanted with the OKa‐C‐1 cell line also showed leukocytosis and hypercalcemia with high serum G‐CSF and PTHrP levels. The culture supernatant of OKa‐C‐1 contained high levels of G‐CSF and PTHrP. Immunohistochemical studies showed the expression of PTHrP in OKa‐C‐1 cells. Reverse transcription polymerase chain reaction revealed the presence of G‐CSF and PTHrP mRNA in this cell line. Dexamethasone treatment inhibited the transcription of G‐CSF and PTHrP genes. This new human squamous carcinoma cell line, OKa‐C‐1, would be useful for studying the mechanism of simultaneous production of G‐CSF and PTHrP and their control in cancer patients with leukocytosis and hypercalcemia.

Keywords: Lung cancer, Squamous cell carcinoma, Cell line, G‐CSF, PTHrP

Full Text

The Full Text of this article is available as a PDF (617.0 KB).

REFERENCES

  • 1. ) Saito , K. , Kuratomi , Y. , Yamamoto , K. , Saito , T. , Kuzuya , T. , Yoshida , S. , Moriyama , S. and Takahashi , A.Primary squamous cell carcinoma of the thyroid associated with marked leukocytosis and hypercalcemia . Cancer , 48 , 2080 – 2083 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Strewler , G. J. , Williams , R. D. and Nissenson , R. A.Human renal carcinoma cells produce hypercalcemia in the nude mouse and a novel protein recognized by parathyroid hormone receptors . J. Clin. Invest. , 71 , 769 – 774 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. ) Kondo , Y. , Sato , K. , Ohkawa , H. , Ueyama , Y. , Okabe , T. , Sato , N. , Asano , S. , Mori , M. , Ohsawa , N. and Kosaka , K.Association of hypercalcemia with tumors producing colony‐stimulating factor(s) . Cancer Res. , 43 , 2368 – 2374 ( 1983. ). [PubMed] [Google Scholar]
  • 4. ) Riddle , P. E. and Dincsoy , H. P.Primary squamous cell carcinoma of the thyroid associated with leukocytosis and hypercalcemia . Arch. Pathol. Lab. Med. , 111 , 373 – 374 ( 1987. ). [PubMed] [Google Scholar]
  • 5. ) Sato , K. , Fujii , Y. , Kakiuchi , T. , Kasono , K. , Imamura , H. , Kondo , Y. , Mano , H. , Okabe , T. , Asano , S. , Takaku , F. , Tsushima , T. and Shizume , K.Paraneoplastic syndrome of hypercalcemia and leukocytosis caused by squamous carcinoma cells (T3M‐1) producing parathyroid hormone related protein, interleukin 1α, and granulocyte colony‐stimulating factor . Cancer Res. , 49 , 4740 – 4746 ( 1989. ). [PubMed] [Google Scholar]
  • 6. ) Rizzoli , R. , Sappino , A. P. and Bonjour , J.‐P.Parathyroid hormone‐related protein and hypercalcemia in pancreatic neuro‐endocrine tumors . Int. J. Cancer , 46 , 394 – 398 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Iguchi , H. , Katakami , H. , Ichinose , Y. , Nishi , Y. , Tanaka , S. , Hara , N. , Ohta , M. , Haji , M. and Nawata , H.A case of squamous cell lung carcinoma with high concentration of parathyroid hormone‐related peptide in serum and pleural effusion presenting hypercalcemia . Jpn. J. Cancer Res. , 84 , 419 – 424 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. ) Matsuoka , S. , Miura , Y. , Kachi , T. , Hattori , Y. , Ohno , J. , Oda , K. , Nagamura , Y. and Oiso , Y.Humoral hypercalcemia of malignancy associated with parathyroid hormone‐related protein producing transitional cell carcinoma of the ureter . Intern. Med. , 33 , 107 – 109 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Yoneda , T. , Nishimura , R. , Kato , I. , Ohmae , M. , Takita , M. and Sakuda , M.Frequency of the hypercalcemia‐leukocytosis syndrome in oral malignancies . Cancer , 68 , 617 – 622 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Asano , S. , Urabe , A. , Okabe , T. , Sato , N. , Kondo , Y. , Ueyama , Y. , Chiba , S. , Ohsawa , N. and Kosaka , K.Demonstration of granulopoietic factor(s) in the plasma of nude mice transplanted with a human lung cancer and in the tumor tissue . Blood , 49 , 845 – 852 ( 1977. ). [PubMed] [Google Scholar]
  • 11. ) Okabe , T. , Sato , N. , Kondo , Y. , Asano , S. , Ohsawa , N. , Kosaka , K. and Ueyama , Y.Establishment and characterization of a human cancer cell line that produces human colony‐stimulating factor . Cancer Res. , 38 , 3910 – 3917 ( 1978. ). [PubMed] [Google Scholar]
  • 12. ) Suva , L. J. , Winslow , G. A. , Wettenhall , R. E. H. , Hammonds , R. G. , Moseley , J. M. , Diefenbach‐Jagger , H. , Rodda , C. P. , Kemp , B. E. , Rodriguez , H. , Chen , E. Y. , Hudson , P. J. , Martin , T. J. and Wood , W. I.A parathyroid hormone‐related protein implicated in malignant hypercalcemia: cloning and expression . Science , 237 , 893 – 896 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Mangin , M. , Webb , A. C. , Dreyer , B. E. , Posillico , J. T. , Ikeda , K. , Weir , E. , Stewart , A. F. , Bander , N. H. , Milstone , L. , Barton , D. E. , Francke , U. and Broadus , A. E.Identification of a cDNA encoding a parathyroid hormone‐like peptide from a human tumor associated with humoral hypercalcemia of malignancy . Proc. Natl. Acad. Sci. USA , 85 , 597 – 601 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. ) Burtis , W. J. , Brady , T. G. , Orloff , J. J. , Ersbak , J. B. , Warrell , R. P. , Olson , B. R. , Wu , T. L. , Mitnick , M. E. , Broadus , A. E. and Stewart , A. F.Immunochemical characterization of circulating parathyroid hormone‐related protein in patients with humoral hypercalcemia of cancer . N. Engl. J. Med. , 322 , 1106 – 1112 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Ueyama , Y. and Tamaoki , N.Leukocytosis in nude mice into which human tumors are transplanted, in vivo screening system for hematopoietic growth factors in human tumors . In “ Hematopoietic Growth Factors: Molecular Biology to Clinical Applications of rG‐CSF .” ed. , Quesenberry P. J. , Asano S. and Saito S. , pp. 27 – 53 ( 1991. ). Excerpta Medica; , Tokyo . [Google Scholar]
  • 16. ) Ellison , M. , Woodhouse , D. , Hillyard , C. , Dowsett , M. , Coombes , R. C. , Gilby , E. D. , Greenberg , P. B. and Neville , A. M.Immunoreactive calcitonin production by human lung carcinoma cells in culture . Br. J. Cancer , 32 , 373 – 379 ( 1975. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. ) Rodan , S. B. , Wesolowski , G. , Ianacone , J. , Thiede , M. A. and Rodan , G. A.Production of parathyroid hormone‐like peptide in a human osteosarcoma cell line: stimulation by phorbol esters and epidermal growth factor . J. Endocrinol , 122 , 219 – 227 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Ichinose , Y. , Iguchi , H. , Ohta , M. and Katakami , H.Establishment of lung cancer cell line producing parathyroid hormone‐related protein . Cancer Lett. , 74 , 119 – 124 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Kinoshita , T. , Imamura , J. , Nagai , H. and Shimotono , K.Quantification of gene expression over a wide range by the polymerase chain reaction . Ann. Biochem. , 206 , 231 – 235 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Metcalf , D. and Nicola , N. A.Proliferative effects of purified granulocyte colony‐stimulating factor (G‐CSF) on normal mouse hemopoietic cells . J. Cell. Physiol. , 116 , 198 – 206 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Nagata , S. , Tsuchiya , M. , Asano , S. , Kaziro , Y. , Yamazaki , T. , Yamamoto , O. , Hirata , Y. , Kubota , N. , Oheda , M. , Nomura , H. and Ono , M.Molecular cloning and expression of cDNA for human granulocyte colony‐stimulating factor . Nature , 319 , 415 – 418 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Demetri , G. D. and Griffin , J. D.Granulocyte colony‐stimulating factor and its receptor . Blood , 78 , 2791 – 2808 ( 1991. ). [PubMed] [Google Scholar]
  • 23. ) Moseley , J. M. , Kubota , M. , Diefenbach‐Jagger , H. , Wettenhall , R. E. H. , Kemp , B. E. , Suva , L. J. , Rodda , C. P. , Ebeling , P. R. , Hudson , P. J. , Zajac , J. D. and Martin , T. J.Parathyroid hormone‐related protein purified from a human lung cancer cell line . Proc. Natl. Acad. Sci. USA , 84 , 5048 – 5052 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Ikeda , K. , Mangin , M. , Dreyer , B. E. , Webb , A. C. , Posillico , J. T. , Stewart , A. F. , Bander , N. H. , Weir , E. C. , Insogna , K. L. and Broadus , A. E.Identification of transcripts encoding a parathyroid hormone‐like peptide in messenger RNAs from a variety of human and animal tumors associated with humoral hypercalcemia of malignancy . J. Clin. Invest. , 81 , 2010 – 2014 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Enomoto , T. , Sugawa , H. , Inoue , D. , Miyamoto , M. , Kosugi , S. , Takahashi , T. , Kitamura , N. , Yamamoto , I. , Konishi , J. , Mori , T. and Imura , H.Establishment of a human undifferentiated thyroid cancer cell line producing several growth factors and cytokines . Cancer , 65 , 1971 – 1979 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Ikeda , K. , Lu , C. , Weir , E. C. , Mangin , M. and Broadus , A. E.Transcriptional regulation of the parathyroid hormone‐related peptide gene by glucocorticoids and vitamin D in a human C‐cell line . J. Biol. Chem. , 264 , 15743 – 15746 ( 1989. ). [PubMed] [Google Scholar]
  • 27. ) Merryman , J. I. , Capen , C. C. , McCauley , L. K. , Werkmeister , J. R. , Suter , M. M. and Rosol , T. J.Regulation of parathyroid hormone‐related protein production by a squamous carcinoma cell line in vitro . Lab. Invest. , 69 , 347 – 354 ( 1993. ). [PubMed] [Google Scholar]
  • 28. ) Binstock , M. L. and Mundy , G. R.Effect of calcitonin and glucocorticoids in combination on the hypercalcemia of malignancy . Ann. Intern. Med. , 93 , 269 – 272 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Merendino , J. J. , Insogna , K. L. , Milstone , L. M. , Broadus , A. E. and Stewart , A. F.A parathyroid hormone‐like protein from cultured human keratinocytes . Science , 231 , 388 – 390 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Fried , R. M. , Voelkel , E. F. , Rice , R. H. , Levine , L. and Tashjian , A. H.Evidence for multiple bone resorption‐stimulating factors produced by normal human keratinocytes in culture . Endocrinology , 122 , 2467 – 2475 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Chodakewitz , J. A. , Kupper , T. S. and Coleman , D. L.Keratinocyte‐derived granulocyte/macrophage colony‐stimulating factor induces DNA synthesis by peritoneal macrophages . J. Immunol. , 140 , 832 – 836 ( 1988. ). [PubMed] [Google Scholar]
  • 32. ) Demetri , G. D. , Zenzie , B. W. , Rheinwald , J. G. and Griffin , J. D.Expression of colony‐stimulating factor genes by normal human mesothelial cells and human malignant mesothelioma cells lines in vitro . Blood , 74 , 940 – 946 ( 1989. ). [PubMed] [Google Scholar]
  • 33. ) Demetri , G. D. , Ernst , T. J. , Pratt , E. S. , II , Zenzie , B. W. , Rheinwald , J. G. and Griffin , J. D.Expression of ras oncogenes in cultured human cells alters the transcriptional and posttranscriptional regulation of cytokine genes . J. Clin. Invest. , 86 , 1261 – 1269 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. ) Li , X. and Drucker , D. J.Parathyroid hormone‐related peptide is a downstream target for ras and src activation . J. Biol. Chem. , 269 , 6263 – 6266 ( 1994. ). [PubMed] [Google Scholar]
  • 35. ) Simmers , R. N. , Webber , L. M. , Shannon , M. F. , Garson , O. M. , Wong , G. , Vadas , M. A. and Sutherland , G. R.Localization of the G‐CSF gene on chromosome 17 proximal to the breakpoint in the t(15;17) in acute promyelocytic leukemia . Blood , 70 , 330 – 332 ( 1987. ). [PubMed] [Google Scholar]
  • 36. ) Chang , J. M. , Metcalf , D. , Gonda , T. J. and Johnson , G. R.Long‐term exposure to retrovirally expressed granulocyte‐colony‐stimulating factor induces a nonneoplastic granulocytic and progenitor cell hyperplasia without tissue damage in mice . J. Clin. Invest. , 84 , 1488 – 1496 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37. ) Keller , P. and Smalling , R.Granulocyte colony stimulating factor: animal studies for risk assessment . Int. Rev. Exp. Pathol. , 34A , 173 – 188 ( 1993. ). [PubMed] [Google Scholar]
  • 38. ) Vernant , J. P. , Brun , B. , Mannoni , P. and Dreyfus , B.Respiratory distress of hyperleukocytic granulocytic leukemias . Cancer , 44 , 264 – 268 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 39. ) Lester , T. J. , Johnson , J. W. and Cuttner , J.Pulmonary leukostasis as the single worst prognostic factor in patients with acute myelocytic leukemia and hyperleukocytosis . Am. J. Med. , 79 , 43 – 48 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 40. ) van Buchem , M. A. , Colly , L. P. , Hogendoorn , P. C. W. , Kluin , P. M. and Willemze , R.Experimental myelocytic leukemia in the brown‐Norway rat as a model for pulmonary leukostasis . Am. J. Pathol. , 138 , 777 – 780 ( 1991. ). [PMC free article] [PubMed] [Google Scholar]
  • 41. ) Vosburgh , E.Pulmonary leukostasis secondary to all‐trans retinoic acid in the treatment of acute promyelocytic leukemia in first relapse . Leukemia , 6 , 608 – 610 ( 1992. ). [PubMed] [Google Scholar]
  • 42. ) Tallman , M. S. and Kwaan , H. C.Reassessing the hemostatic disorder associated with acute promyelocytic leukemia . Blood , 79 , 543 – 553 ( 1992. ). [PubMed] [Google Scholar]
  • 43. ) Frankel , S. R. , Eardley , A. , Lauwers , G. , Weiss , M. and Warrell , R. P. , Jr.The “retinoic acid syndrome” in acute promyelocytic leukemia . Ann. Intern. Med. , 117 , 292 – 296 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 44. ) Oeda , E. , Shinohara , K. , Kamei , S. , Nomiyama , J. and Inoue , H.Capillary leak syndrome likely the result of granulocyte colony‐stimulating factor after high‐dose chemotherapy . Intern. Med. , 33 , 115 – 119 ( 1994. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES