Regulative effect of cyclooxygenase-2 3'-UTR in HT29 colon cancer cells / 中华消化外科杂志

ABSTRACT

Objective To investigate the regulative effect of cyclooxygenase-2 (COX-2) 3'-UTR in HT29 colon cancer cells. Methods Total RNA was extracted from HT29 colon cancer cells and used as a template to amplify COX-2 gene by reverse transcription polymerase chain reaction. Using PCR technique to construct a series of luciferase reporter gene expression vectors containing various regions of the 3'-UTR of COX-2 (including the whole gene region, gene region between 1 to 156 bp, gene region between 1 to 347 bp, gene region between 1 to 1006 bp, gene region between 156 to 347 bp and gene region between 157 to 2217 bp). These reporter gene expression vectors and pRL-SV40 were co-transfected in HT29 colon cancer cells by Lipofectamine 2000. The relative luciferase activity of the HT29 colon cancer cells was tested. All data were analyzed via t test. Results The luciferase activity was reduced by 70.4%, 37.4%, 64.8% and 24.2% in HT29 colon cancer cells transfected with the whole COX-2 gene region, gene region between 1 to 156 bp, gene region between 1 to 347 bp and gene region between 156 to 347 bp, respectively (t = 6.13, 7.73, 9.75, 3.92, P < 0.05). No obvious changes of luciferase activity were observed in HT29 colon cancer cells transfected with gene regions between 1 to 1006 bp and between 157 to 2217 bp (t = 0.13, 0.01, P > 0.05). Conclusions A region between 156-347 bp in the 3'-UTR of COX-2 has been found which can down-regulate the expression of COX-2 with the cooperation of the ARE element. The 3'-UTR of COX-2 contains several control elements that regulate the expression of COX-2 in colon cancer cells.