Reciprocal regulation of cyclooxygenase-2 and 15-hydroxyprostaglandin dehydrogenase expression in A549 human lung adenocarcinoma cells.
Human lung adenocarcinoma cells, A549, possess the capacity of expressing both cyclooxygenase-2 (COX-2) and NAD+-linked 15-hydroxyprostaglandin dehydrogenase (15-PGDH). Resting cells express little COX-2 but significant levels of 15-PGDH. Interleukin (IL) 1beta, tumor necrosis factor-alpha (TNF-alpha) or phorbol ester [phorbol 12-myristate 13-acetate (PMA)] induced the expression of COX-2, as revealed by western blot analysis. Combination of PMA and IL-1beta or TNF-alpha induced synergistically the expression of COX-2. Interestingly, cytokines and cytokine plus PMA-induced expression of COX-2 were accompanied by a downregulation of 15-PGDH. This was evident from both the western blot analysis and activity assay of 15-PGDH. It appears that the higher the expression of COX-2 was induced, the lower the expression of 15-PGDH was found. This was further supported by the observation that overexpression of COX-2 but not COX-1 by adenovirus-mediated approach led to a decrease in 15-PGDH expression, indicating the specificity of COX-2. Furthermore, downregulation of the IL-1beta-induced expression of COX-2 by silencing RNA (siRNA) approach resulted in an increase in the expression of 15-PGDH by COX-2-siRNA but not by COX-1-siRNA, indicating that it was indeed the expression of COX-2 attenuating the expression of 15-PGDH. The IL-1beta-induced reduction of the expression of 15-PGDH was shown not to be mediated by COX-2-derived products since the presence of COX-2 inhibitors did not block the attenuation of the expression of 15-PGDH. Exogenous PGE2 also did not induce the reduction of the expression of 15-PGDH. However, overexpression of 15-PGDH by transfection with recombinant plasmid encoding 15-PGDH or adenovirus-mediated approach attenuated IL-1beta-induced expression of COX-2. On the contrary, downregulation of 15-PGDH expression by 15-PGDH-siRNA or 15-PGDH-antisense approach resulted in an increase in IL-1beta-induced expression of COX-2 but not that of COX-1. In fact, it was further observed that A549 clones expressing different degrees of 15-PGDH showed also different levels of COX-2 expression after IL-1beta induction. The levels of IL-1beta-induced COX-2 expression appeared to correlate inversely with those of 15-PGDH expression in the cells. These results support the contention that COX-2 and 15-PGDH are regulated reciprocally in A549 cells.
Tong
Min
M
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky Lexington, KY 40536-0082, USA.
Ding
Yunfei
Y
Tai
Hsin-Hsiung
HH
D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology; D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000096:biosynthesis*; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D006911:Hydroxyprostaglandin Dehydrogenases / Q000096:biosynthesis*; D007124:Immunoenzyme Techniques; D008175:Lung Neoplasms / Q000201:enzymology* / Q000473:pathology; D034741:RNA, Small Interfering / Q000378:metabolism
D034741:RNA, Small Interfering; D006911:Hydroxyprostaglandin Dehydrogenases; C035859:15-hydroxyprostaglandin dehydrogenase; D051546:Cyclooxygenase 2