TITLE

IMMEDIATE AND EARLY GENE RESPONSE TO IN VITRO ACID EXPOSURE IN BARRETT'S ADENOCARCINOMA CELL LINE

AUTHOR(S)
Morgan, C.; Alazawi, W.; Sirieix, P.; Freeman, T.; Coleman, N.; Fitzgerald, R.
PUB. DATE
April 2003
SOURCE
Gut;Apr2003 Supplement 1, Vol. 52, pA44
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Introduction: Acid, a principal component of refluxate, may contribute to the neoplastic progression of Barrett's oesophagus. Previously published data have demonstrated that brief acid exposure in vivo and in vitro increases cell proliferation. The mechanisms underlying the hyperproliferative response are not well elucidated but may include alterations in Na/H exchanger activity and MAPK signalling pathways. Aim: To ascertain the effects of acid exposure on gene expression in a Barrett's adenocarcinoma cell line (SEG-1) using expression micro arrays and RT-PCR. Methods: SEG-1 cells were grown to 60% confluency and exposed to either acidified DMEM at pH 3.5 (0.1M HCI) or pH 7.4 (control) for 20 minutes followed by neutralisation of the medium for up to 10 hours. Total RNA was extracted before acid exposure and over a 10 hour time course (0.5, 2, 4, 6, 8, and 10h) and hybridised to an Affymetrix human U133A oligonucleotide array. Data were analysed using the Affymetrix statistical expression algorithms. Only alterations in gene expression >2, <-2 were taken as significant and a subset of interest were validated by RT-PCR. Results: An up-regulation of genes associated with proliferation (PCNA, FGFR3 and VEGFC) and a down-regulation of genes associated with apoptosis (caspase-9, GADD45A) were shown throughout all time points. DUSP2 and 8, which are involved in the inactivation of the MAPK pathway were down regulated throughout the time course. At specific time points the following cell cycle regulatory genes were significantly altered: E2F (up at 0.5h), Rb binding protein 2 homolog 1(down at 0.5, 2h), Cyclin E2 (up at 2, 6, 8h) Cyclin E1 (up at 4, 6h) and Cyclins D1 and A1 (up at 8h). Cyclo-oxygenases were unaffected. Conclusion: Suppression of apoptosis via the p53 pathway, stimulation of proliferation via the MAPK pathway and alterations in cell cycle components may be involved in the proliferative response to an acid pulse. This study provides candidate genes...
ACCESSION #
9747564

 

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