Vasoactive intestinal peptide: A potent stimulator of adenosine 3′:5′-cyclic monophosphate accumulation in gut carcinoma cell lines in culture

Abstract

Vasoactive intestinal peptide (VIP) is a potent and efficient stimulator of adenosine 3':5'-cyclic monophosphate (cAMP) accumulation in a human colon carcinoma cell line, HT 29. cAMP accumulation is sensitive to a concentration of VIP as low as 3 X 10-12 M. Maximum VIP-induced cAMP levels were observed with 10-9 M VIP and are about 200 times above the basal levels. Half-maximum cAMP production was obtained at 3 X 10-1°M VIP. 125I-Labeled VIP was found to bind to HT 29 cells; this binding was competitively inhibited by concentrations of unlabeled VIP between 10-10 and 10-7 M. Half-maximum inhibition of binding was observed with 2 X 10-9 M VIP. For experimental studies, 3-4 X 106 cells were transfered into 150-cm2 flasks in 30 ml of culture medium. The medium was changed on day 4, and 3 days later, cells were collected with the buffer described above, without trypsin. Under such conditions, the cells were in exponential growth phase. The average yield was 30 X 106 cells per flask. The cells were washed three times in Krebs-Ringer phosphate buffer, pH 7.4, containing 2%

Photoaffinity Labeling Demonstrates Physical Contact between Vasoactive Intestinal Peptide and the N-terminal Ectodomain of the Human VPAC1 Receptor

“…In consonance with its ubiquitous distribution, VIP controls a large array of biological functions (1)(2)(3). A non-exhaustive list includes exocrine secretions, release of hormones, neuronal excitation, relaxation of muscles, metabolism, immune function, inflammatory response, growth control of fetuses, embryonic brain development, and tumor cell biology (1)(2)(3)(4)(5). VIP action on its numerous target cells is mediated by two serpentine G protein-coupled receptors referred to as VPAC1 and VPAC2 (6).…”

Expression of glucagon‐like peptide 1 receptor in a murine C cell line Regulation of calcitonin gene by glucagon‐like peptide 1

“…trol) or presence of different peptides was measured by RIA [16] from cells treated as previously described [17]. Porcine glucagon was obtained from Novo Research Institute (Bagsvaerdt, Denmark), and porcine oxyntomodulin was a gift from Dr. D. Bataille (INSERM Unit 376, Montpellier).…”

Covalent cross‐linking of vasoactive intestinal peptide (VIP) to its receptor in intact colonic adenocarcinoma cells in culture (HT 29)

“…This cell has conserved several characteristics of the native tissue, in particular receptor sites with high affinity for VIP, a very large increase in cAMP level in response to receptor site occupancy [14] and subsequent parallel activation of CAMP-dependent protein kinases and cAMP phosphodiesterase [15, 161. In the present work we have used the cleavable cross-linking reagent dithiobis(succinimidylpropionate) (DTSP) to link covalently radioactive monoiodinated VIP to VIP receptors in HT 29 cell monolayers. Our results demonstrated the existence of a major polypeptide of M , = 64000, which represents the unique class of high-affinity binding sites in intact HT -1 pM).…”