Document Detail


Quantification of elastase-like activity in 13 human cancer cell lines and in an immortalized human epithelial cell line by RP-HPLC.
MedLine Citation:
PMID:  12817479     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A sensitive and specific RP-HPLC assay was developed to measure the levels of polymorphonuclear elastase (PMN-E) activity in growing cell cultures. By combining a pre-incubation of the cells with a relatively non-toxic, PMN-E-specific inhibitor, MeOSuc-Ala-Ala-Pro-Val-chloromethylketone (MAAPVCK), the p-nitroaniline formed by the hydrolysis of the substrate MeOSuc-Ala-Ala-Pro-Val-p-NA by PMN-E is quantified. Elastase-like activity was measured in 14 human cells lines: 13 cancer cell lines (HL-60, U-937, A-427, LCLC-103H, YAPC, DAN-G, PA-TU-8902, KYSE-70, -510, -520, 5637, SISO and MCF-7) and one immortalized epithelial cell line (hTert-RPE1). Activity was detected in all lines; the lowest was found in hTert-RPE1 cells while the highest was detected in a pancreas adenocarcinoma line (PA-TU-8902). When the results were normalized according to cell volume instead of cell number, the leukemia line HL-60 had the highest activity and PA-TU-8902 ranked second. A 1 h pre-incubation with 9.0 microM of the irreversible PMN-E inhibitor MAAPVCK led to varying degrees of enzyme inhibition depending on the cell line; the strongest inhibition was observed with the PA-TU-8902 pancreatic cancer cell line (90% inhibition) while the weakest was seen with the A-427 lung cancer cell line (52%). These results indicate that PA-TU-8902 is a suitable in vitro model for testing the efficacy of PMN-E-activated prodrugs of antitumor agents.
Authors:
Karin Achilles; Patrick J Bednarski
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biological chemistry     Volume:  384     ISSN:  1431-6730     ISO Abbreviation:  Biol. Chem.     Publication Date:  2003 May 
Date Detail:
Created Date:  2003-06-23     Completed Date:  2004-02-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9700112     Medline TA:  Biol Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  817-24     Citation Subset:  IM    
Affiliation:
Institute of Pharmacy, Ernst-Moritz-Arndt University, D-17487 Greifswald, Germany.
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MeSH Terms
Descriptor/Qualifier:
Aniline Compounds / analysis
Cell Division / drug effects,  physiology
Cell Line
Cell Line, Tumor
Cell Size
Chromatography, High Pressure Liquid / methods
Enzyme Inhibitors / pharmacology
Epithelial Cells / cytology,  metabolism
Humans
Leukocyte Elastase / antagonists & inhibitors,  metabolism*
Neoplasms / enzymology*
Chemical
Reg. No./Substance:
0/Aniline Compounds; 0/Enzyme Inhibitors; 100-01-6/4-nitroaniline; EC 3.4.21.37/Leukocyte Elastase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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