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

Randy S. Haun, Margery C. Beinfeld, Bernard A. Roos, Jack E. Dixon

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

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 (US)
Pages (from-to)850-856
Number of pages7
JournalEndocrinology
Volume125
Issue number2
DOIs
StatePublished - Aug 1989

ASJC Scopus subject areas

  • Endocrinology

Fingerprint Dive into the research topics of 'Establishment of a cholecystokinin-producing rat medullary thyroid carcinoma cell line'. Together they form a unique fingerprint.

Cite this