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Production of cytokines by oral cancer cells cultured in a protein-free system

Research Project

Project/Area Number 02807180
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 外科・放射線系歯学
Research InstitutionTokushima University (1991)
Tokyo Medical and Dental University (1990)

Principal Investigator

RIKIMARU Koichi  Tokushima Univ., Univ. Dent. Hosp., Assist. Prof., 歯学部附属病院, 講師 (40220800)

Co-Investigator(Kenkyū-buntansha) 堀越 勝  東京医科歯科大学, 歯学部, 文部教官助手 (00014283)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1991: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1990: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsOral Cancer / Protein-Free Medium / Protein-Free Culture / Squamous Cell Carcinoma / Cytokine / Growth Factor
Research Abstract

A protein-free cell culture system for human squamous cell carcinoma of the oral cavity has been established. Using this protein-free culture system, it was revealed that human gingival carcinoma derived cell line Ca9-22PF, which has been cultured with a protein-free synthetic medium PF86-1, produced IL-1 and PDGF-AA constitutively. However, it seems to be very hard to screen out the many kinds of cytokines produced by oral cancer cells only using a proteinfree culture system. Therefore, in this study we tried to detect the expression of gene for cytokines by the polymerase chain reaction (PCR) method.
Cytoplasmic RNA was prepared from ZAPF cells, which was established from a metastatic lymph node of a palatal cancer and cultured with PF86-1, using Nonidet P-40. Further purification of mRNA was carried out by loading it on a oligo-dT cellulose column. Single strand cDNAs were synthesized with MO-MLV reverse transcriptase from this MRNA used as a template, then targeted DNA for cytokines, G-CSF, M-CSF, GM-CSF, IL-3 and IL-6 respectively, was amplified, if it had existed in the sample, by PCR method and separated by agarose gel electrophoresis (AGE).
As results, no predicted band was observed on AGE, when PCR was performed for G-CSF, IL-3 and IL-6. On the other hand, in the case of G-CSF 630 base pairs (bp) band, which was predicted from the sequence of cDNA, was visualized on AGE. In the case of M-CSF, a predicted 450 bp band was detected and it was separated to two bands of 150bp and 300bp prospected from the cDNA sequence after the digestion with Mung Bean Nulease and Stu I restriction endonuclease.
These results show that ZAPF expresses at least two cytokine genes, GM-CSF and M-CSF, however it will be necessary to investigate whether ZAPF produce GMCSF and M-CSF at biologically responsible concentration or not using a proteinfree culture system.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] 立川 敬子: "口腔扁平上皮癌細胞の産生する骨吸収促進因子に関する研究" 日本口腔外科学会雑誌. 36. 964-978 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kaechong Lee: "Epidermal Growth Factor Stimulates the Anchorage-Independent Growth of Human Squamous Cell Carcinomas Overexpressing Its Receptors." Biochemical and Biophysical Research Communications. 168. 905-911 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Koichi Rikimaru: "Growth of the Malignant and Nonmalignant Human Squamous Cells in a Protein-Free Defined Medium." In Vitro Cellular and Developmental Biology. 26. 849-856 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 力丸 浩一: "ヌ-ドマウス移植扁平上皮癌の増殖に対するヒトEGFの影響" 日本口腔外科学会雑誌. 38. 23-29 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Koichi Rikimaru: "Gene Amplification and Overexpression of Epidermal Growth Factor Receptor in Squamous Cell Carcinoma of the Head and Neck." Head and Neck. 14. 8-13 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Noriko Tachikawa: "Bone resorbing factors produced by oral squamous cell carcinoma." Jpn. J. Oral. Maxillofac. Surg.36. 964-978 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kaecong Lee: "Epidermal growth factor stimulates the anchorage-independent growth of human squamous cell carcinomas overexpressing its receptor." Biocem. Biophys. Res. Commun.168. 905-911 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Koichi Rikimaru: "Growth of the malignant and nonmalignant human squamous cells in a protein-free defined medium." In Vitro Cell. Dev. Biol.26. 849-856 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Koichi Rikimaru: "Effects of human EGF on the growth of human squamous cell carcinomas in athymic mice." Jpn. J. Oral. Maxillofac. Surg.38. 23-29 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Koichi Rikimaru: "Gene amplification and overexpression of epidermal growth receptor in squamous cell carcinoma of the head and neck." Head and Neck. 14. 8-13 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 立川 敬子: "口腔扁平上皮癌細胞の産生する骨吸収促進因子に関する研究" 日本口腔外科学会雑誌. 36. 964-978 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kaechong Lee: "Epidermal Growth Factor Stimulates the AnchorageーIndependent Growth of Human Squamous Cell Carcinomas Overexpressing Its Receptors." Biochemical and Biophysical Research Communications. 168. 905-911 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Koichi Rikimaru: "Growth of the Malignant and Nonmalignant Human Squamous Cells in a ProteinーFree Defined Medium." In Vitro Cellular and Developmental Biology. 26. 849-856 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] 力丸 浩一: "ヌ-ドマウス移植扁平上皮癌の増殖に対するヒトEGFの影響" 日本口腔外科学会雑誌. 38. 23-29 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Koichi Rikimaru: "Gene Amplification and Overexpression of Epidermal Growth Factor Receptor in Squamous Cell Carcinoma of the Head and Neck." Head and Neck. 14. 8-13 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 立川 敬子: "口腔扁平上皮癌細胞の産生する骨吸収促進因子に関する研究" 日本口腔外科学会雑誌. 36. 964-978 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kaechoong Lee: "Epidermal Growth Factor Stimulates the Anchorageーindependent Growth of Human Squamous Cell Carcinomas Overexprossing Its Receptors" Biochemical and Biophysical Research Commanications. 168. 905-911 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Koichi Rikimaru: "Grooth of the Malignant and Nonmalignant Human Squamous Cells in A Proteinーfree Defined Medium" IN VITRO Cellular and Developmental Biology. 26. 849-856 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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