학술논문
Preliminary detection of the anti-tumour activity of indoline-2,3-dione derivative DH-12a targeting aminopeptidase N
Document Type
Report
Author
Source
Molecular Medicine Reports. November 1, 2014, Vol. 10 Issue 5, p2681, 8 p.
Subject
Language
English
ISSN
1791-2997
Abstract
Introduction Aminopeptidase N (APN; EC 3.4.11.2; CD13), also termed CD13, is a type 2 transmembrane zinc-dependent metallopeptidase of the superfamily of gluzincins (1,2). APN is expressed in numerous cells and [...]
Aminopeptidase N (APN) is important in tumour processes. The present study detected the anti-tumour activity of the novel APN inhibitor DH-12a, which is an indoline-2,3-dione derivative. In the present study, Bestatin, a clinical APN inhibitor was used as a positive control. The expression of APN in the ES-2 and 3AO cell lines were assessed using flow cytometry and the drug inhibition constants of DH-12a ([K.sub.i] = 13.15 µM) and Bestatin ([K.sub.i] = 16.57 µM) were assessed using a double reciprocal method of competitive inhibition. The in vitro effects of DH-12a on cell proliferation were assessed using a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay on human cell lines of ES-2 ([IC.sub.50] = 43.8 µM), A549 (inhibition rate = 41.5% at 160 µM DH-12a), HL60 (inhibition rate = 47.83% at 160 µM DH-12a) and 3AO (IC50 = 70.2 µM). The inhibition rates were consistently higher than those of Bestatin. The effects of DH-12a on cell migration (inhibition rates in ES-2 cells and 3AO cells were 56.4 and 76.5%, respectively at 15 µM) and invasion (inhibition rates in ES-2 cells and 3AO cells were 75.6 and 66.5%, respectively at 15 µM) were assessed using transwell plates. The in vivo effects of DH-12a on tumour proliferation and lung tumour metastasis were determined using an H22 xenograft mice model, where DH-12a was administered in combination with genotoxic 5-fluorouracil. The anti-tumour activities of DH-12a in vivo were also greater than those of Bestatin. In conclusion, the in vitro effects of DH-12a on tumour proliferation, migration and invasion were consistent with the in vivo effects. In addition, DH-12a exhibited greater anti-tumour properties compared with Bestatin. Key words: aminopeptidase N, indoline-2,3-dione derivative, migration, invasion
Aminopeptidase N (APN) is important in tumour processes. The present study detected the anti-tumour activity of the novel APN inhibitor DH-12a, which is an indoline-2,3-dione derivative. In the present study, Bestatin, a clinical APN inhibitor was used as a positive control. The expression of APN in the ES-2 and 3AO cell lines were assessed using flow cytometry and the drug inhibition constants of DH-12a ([K.sub.i] = 13.15 µM) and Bestatin ([K.sub.i] = 16.57 µM) were assessed using a double reciprocal method of competitive inhibition. The in vitro effects of DH-12a on cell proliferation were assessed using a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay on human cell lines of ES-2 ([IC.sub.50] = 43.8 µM), A549 (inhibition rate = 41.5% at 160 µM DH-12a), HL60 (inhibition rate = 47.83% at 160 µM DH-12a) and 3AO (IC50 = 70.2 µM). The inhibition rates were consistently higher than those of Bestatin. The effects of DH-12a on cell migration (inhibition rates in ES-2 cells and 3AO cells were 56.4 and 76.5%, respectively at 15 µM) and invasion (inhibition rates in ES-2 cells and 3AO cells were 75.6 and 66.5%, respectively at 15 µM) were assessed using transwell plates. The in vivo effects of DH-12a on tumour proliferation and lung tumour metastasis were determined using an H22 xenograft mice model, where DH-12a was administered in combination with genotoxic 5-fluorouracil. The anti-tumour activities of DH-12a in vivo were also greater than those of Bestatin. In conclusion, the in vitro effects of DH-12a on tumour proliferation, migration and invasion were consistent with the in vivo effects. In addition, DH-12a exhibited greater anti-tumour properties compared with Bestatin. Key words: aminopeptidase N, indoline-2,3-dione derivative, migration, invasion