Comparison of Absolute Expression and Turnover Number of COX-1 and COX-2 in Human and Rodent Cells and Tissues
© 2022 Li et al..
Objective: We aim to quantify the absolute protein expression of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in various cells and tissues to determine the relative contribution of COX-1 and COX-2 to PGE2 production.
Methods: An LC-MS method was developed and validated, then used for quantifying the absolute amounts of COX-1 and COX-2 in recombinant human COX-1 and COX-2, lysates from different cells, tissue microsomes of rodents and humans, Pirc rat colonic polyps, and biopsy specimens from squamous cell carcinoma (SCC) patients. The COX-1 and COX-2 turnover numbers were subsequently calculated based on apparent formation rates of PGE2.
Results: A robust LC-MS method for quantification of COX-1 and COX-2 was developed and validated and then used to calculate their apparent turnover numbers. The results showed that COX-1 expression levels were much higher than that of COX-2 in all the tested tissues including the colonic epithelium of F344 (28-fold) and Pirc rats (20-fold), colonic polyps of Pirc rats (8-fold), and biopsy specimens of SCC patients (11-17-fold). In addition, both COX-1 and COX-2 were higher in polyps when compared to adjacent mucosa of Pirc rats. The turnover number of recombinant human COX-2 was 14-fold higher than that of recombinant human COX-1. LPS stimulation increased COX-2 protein expression in three cell lines (Raw 264.7, SCC9 and EOMA) as expected but unexpectedly increased COX-1 protein expression (13.8-fold) in EOMA cells.
Conclusion: In human oral cancer tissues and cells as well as Pirc rat colon, COX-1 plays an unexpectedly but more important role than COX-2 in abnormal PGE2 production since COX-1 expression is much higher than COX-2. In addition, COX-1 expression levels are inducible in cells, and higher in polyps than surrounding mucosa in Pirc rat colon. These results indicate that targeted suppression of local COX-1 should be considered to reduce colon-specific PGE2-mediated inflammation.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:15 |
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| Enthalten in: |
Journal of inflammation research - 15(2022) vom: 15., Seite 4435-4447 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Li, Li [VerfasserIn] |
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| Links: |
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| Themen: |
Absolute quantification |
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| Anmerkungen: |
Date Revised 13.08.2022 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
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| doi: |
10.2147/JIR.S365842 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 100 | 1 | |a Li, Li |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Comparison of Absolute Expression and Turnover Number of COX-1 and COX-2 in Human and Rodent Cells and Tissues |
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| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a © 2022 Li et al. | ||
| 520 | |a Objective: We aim to quantify the absolute protein expression of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in various cells and tissues to determine the relative contribution of COX-1 and COX-2 to PGE2 production | ||
| 520 | |a Methods: An LC-MS method was developed and validated, then used for quantifying the absolute amounts of COX-1 and COX-2 in recombinant human COX-1 and COX-2, lysates from different cells, tissue microsomes of rodents and humans, Pirc rat colonic polyps, and biopsy specimens from squamous cell carcinoma (SCC) patients. The COX-1 and COX-2 turnover numbers were subsequently calculated based on apparent formation rates of PGE2 | ||
| 520 | |a Results: A robust LC-MS method for quantification of COX-1 and COX-2 was developed and validated and then used to calculate their apparent turnover numbers. The results showed that COX-1 expression levels were much higher than that of COX-2 in all the tested tissues including the colonic epithelium of F344 (28-fold) and Pirc rats (20-fold), colonic polyps of Pirc rats (8-fold), and biopsy specimens of SCC patients (11-17-fold). In addition, both COX-1 and COX-2 were higher in polyps when compared to adjacent mucosa of Pirc rats. The turnover number of recombinant human COX-2 was 14-fold higher than that of recombinant human COX-1. LPS stimulation increased COX-2 protein expression in three cell lines (Raw 264.7, SCC9 and EOMA) as expected but unexpectedly increased COX-1 protein expression (13.8-fold) in EOMA cells | ||
| 520 | |a Conclusion: In human oral cancer tissues and cells as well as Pirc rat colon, COX-1 plays an unexpectedly but more important role than COX-2 in abnormal PGE2 production since COX-1 expression is much higher than COX-2. In addition, COX-1 expression levels are inducible in cells, and higher in polyps than surrounding mucosa in Pirc rat colon. These results indicate that targeted suppression of local COX-1 should be considered to reduce colon-specific PGE2-mediated inflammation | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a cyclooxygenase-2 | |
| 650 | 4 | |a inflammation | |
| 650 | 4 | |a liquid chromatography with tandem mass spectrometry | |
| 650 | 4 | |a turnover number | |
| 700 | 1 | |a Sun, Rongjin |e verfasserin |4 aut | |
| 700 | 1 | |a Zenga, Joseph |e verfasserin |4 aut | |
| 700 | 1 | |a Himburg, Heather |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Lu |e verfasserin |4 aut | |
| 700 | 1 | |a Duan, Shengnan |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Jingwen |e verfasserin |4 aut | |
| 700 | 1 | |a Bui, Dinh |e verfasserin |4 aut | |
| 700 | 1 | |a Xie, Zuoxu |e verfasserin |4 aut | |
| 700 | 1 | |a Du, Ting |e verfasserin |4 aut | |
| 700 | 1 | |a Xie, Lijun |e verfasserin |4 aut | |
| 700 | 1 | |a Yin, Taijun |e verfasserin |4 aut | |
| 700 | 1 | |a Wong, Stu |e verfasserin |4 aut | |
| 700 | 1 | |a Gao, Song |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Ming |e verfasserin |4 aut | |
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