Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line

R. S. Haun, M. C. Beinfeld, B. A. Roos, J. E. Dixon

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

A rat medullary thyroid carcinoma, which was previously shown to produce high levels of immunoreactive cholecystokinin (CCK), was used to establish a stable cell line. Transplantable tumors were subjected to four series of alternate in vitro and in vivo passages. Cells were prepared from the fourth series of tumors under serum-free medium conditions that prevent fibroblasts growth. Subcloning of these cells yielded several propagatable clonal cell lines. One cell line with immunoreactive CCK-8 production was selected for further studies. This high CCK cell line, WE4/2, produces and secretes a CCK-immunoreactive product that coelutes with synthetic CCK-8 sulfate during Sephadex chromatography and HPLC. Northern analysis with a rat CCK cDNA revealed that the cultured cells produce a CCK RNA the same size and with the same 5' end as that previously reported for brain and intestines. In addition, a recombinant plasmid containing about 800 basepairs of 5' flanking sequence of the rat CCK gene linked to the coding sequence of the bacterial chloramphenicol acetyltransferase gene elicited a high level of chloramphenicol acetyltransferase activity when transfected into the WE4/2 cell line. Therefore, the WE4/2 cell line provides a model system for studying CCK gene expression and biosynthesis.

Original languageEnglish
Pages (from-to)850-856
Number of pages7
JournalEndocrinology
Volume125
Issue number2
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

Cholecystokinin
Cell Line
Chloramphenicol O-Acetyltransferase
5' Flanking Region
Serum-Free Culture Media
Medullary Thyroid cancer
Genes
Sulfates
Intestines
Chromatography
Cultured Cells
Neoplasms
Plasmids
Complementary DNA
Fibroblasts
High Pressure Liquid Chromatography
RNA
Gene Expression
Brain
Growth

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Haun, R. S., Beinfeld, M. C., Roos, B. A., & Dixon, J. E. (1989). Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line. Endocrinology, 125(2), 850-856.

Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line. / Haun, R. S.; Beinfeld, M. C.; Roos, B. A.; Dixon, J. E.

In: Endocrinology, Vol. 125, No. 2, 01.01.1989, p. 850-856.

Research output: Contribution to journalArticle

Haun, RS, Beinfeld, MC, Roos, BA & Dixon, JE 1989, 'Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line', Endocrinology, vol. 125, no. 2, pp. 850-856.
Haun RS, Beinfeld MC, Roos BA, Dixon JE. Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line. Endocrinology. 1989 Jan 1;125(2):850-856.
Haun, R. S. ; Beinfeld, M. C. ; Roos, B. A. ; Dixon, J. E. / Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line. In: Endocrinology. 1989 ; Vol. 125, No. 2. pp. 850-856.
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