Effects of parathyroid hormone-related peptide on adenosine 3′,5′-monophosphate and ornithine decarboxylase in a human colonic cell line

Dahong Yu, Patricia K. Seitz, Peter Selvanayagam, Srinivasan Rajaraman, Courtney Townsend, Cary W. Cooper

Research output: Contribution to journalArticle

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Abstract

PTH-related peptide (PTHrP) is widely distributed in normal tissues, including the gut, and is considered a potential autocrine or paracrine regulator of cellular growth and differentiation. With this in mind, a human colonic cell line (LoVo) was used to study the effect of PTHrP on ornithine decarboxylase (ODC), because ODC is known to have profound effects on the growth and differentiation of many cell types via stimulation of synthesis of polyamines. cAMP also was measured, because this second messenger has been implicated in the regulation of ODC activity. Nearly confluent LoVo cells, grown in F-12 medium and 10% fetal bovine serum (FBS), were preincubated in 1% FBS for at least 5 h, and then PTHrP-(1-34) was added, and the incubation was continued for up to 6 h. Cell extracts were analyzed for ODC activity by measuring 14CO2 liberated from 14C-labeled ornithine, for cAMP by RIA, and for ODC mRNA by Northern analysis. PTHrP produced dose-related increases in both cAMP (2- to 3-fold) and ODC (3- to 5-fold), with a maximal effect at 0.1-1 μM and an ED50 of 1-10 nM. Comparison of the cAMP and ODC responses to PTHrP showed a strong correlation (r = 0.96; P < 0.001). The effects of 1 μM PTHrP-(1-34) to increase cAMP and ODC were completely inhibited by 10-20 μM of the specific antagonist [Asn10,Leu11]PTHrP-(7-34). PTHrP-(1-34) did not stimulate ODC activity when cells were incubated without FBS. The stimulation of ODC activity by PTHrP-(1-34) was maximal at 2 h, a time at which an increase in ODC mRNA also was evident. PTH-(1-34) and forskolin also stimulated ODC activity, but PTHrP-(67-86) amide was ineffective. The results indicate that the N-terminal portion of the PTHrP molecule can stimulate ODC activity in a human colon cell line and that the effect is probably mediated by cAMP. The results are consistent with the idea that PTHrP may influence cell growth and differentiation in the gut via an effect on polyamine biosynthesis. Since LoVo cells also express PTHrP mRNA, this gastrointestinal cell line may serve as a useful model for studying autocrine regulation of gut cell growth and differentiation by PTHrP.

Original languageEnglish (US)
Pages (from-to)1993-2000
Number of pages8
JournalEndocrinology
Volume130
Issue number4
StatePublished - Apr 1992

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Parathyroid Hormone-Related Protein
Ornithine Decarboxylase
Adenosine
Cell Line
Cell Differentiation
Polyamines
Growth
Messenger RNA
Serum
Ornithine
Second Messenger Systems
Colforsin
Cell Extracts
Parathyroid Hormone
Amides

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Effects of parathyroid hormone-related peptide on adenosine 3′,5′-monophosphate and ornithine decarboxylase in a human colonic cell line. / Yu, Dahong; Seitz, Patricia K.; Selvanayagam, Peter; Rajaraman, Srinivasan; Townsend, Courtney; Cooper, Cary W.

In: Endocrinology, Vol. 130, No. 4, 04.1992, p. 1993-2000.

Research output: Contribution to journalArticle

Yu, Dahong ; Seitz, Patricia K. ; Selvanayagam, Peter ; Rajaraman, Srinivasan ; Townsend, Courtney ; Cooper, Cary W. / Effects of parathyroid hormone-related peptide on adenosine 3′,5′-monophosphate and ornithine decarboxylase in a human colonic cell line. In: Endocrinology. 1992 ; Vol. 130, No. 4. pp. 1993-2000.
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