LYP, a novel bestatin derivative, inhibits cell growth and suppresses APN/CD13 activity in human ovarian carcinoma cells more potently than bestatin
- 291 Downloads
LYP is a bestatin dimethylaminoethyl ester which inhibits aminopeptidase N (APN/CD13). Our goal in this study was to evaluate LYP as a candidate compound for cancer treatment, beginning by studying its inhibitory effects on tumors and then comparing it to bestatin. Experiments were performed on human ovarian carcinoma (OVCA) ES-2 and SKOV-3 cell lines, which have high and low levels of APN/CD13 respectively. LYP effectively inhibited ES-2 cell growth as estimated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and the trypan blue dye-exclusion test. LYP significantly suppressed APN/CD13 activity on the surface of ES-2 cells as measured by quantifying the enzymatic cleavage of the substrate L-leucine-p-nitroanilide. The inhibitory effects of LYP were greater than those of bestatin at the same concentrations. In contrast, LYP was a weak inhibitor of SKOV-3 cell growth, suggesting that LYP may inhibit ES-2 cell growth via suppression of APN/CD13. Inhibition of APN/CD13 expression was also demonstrated with immunofluorescent flow cytometry and Western blot analysis. Inhibitory effects of LYP were confirmed by using a mouse model in which LYP delayed the growth of ES-2 xenografts in mice after 2 weeks of LYP injections. Inhibition of APN/CD13 expression was demonstrated in the ES-2 xenografts using Western blot analysis. The inhibitory effects of LYP on the ES-2 xenografts were stronger than those of bestatin. These results suggest that LYP has a powerful inhibitory effect on the growth of OVCA cells and that the mechanism may be via a decrease in the expression of APN/CD13.
KeywordsAminopeptidase N (APN/CD13) Ovarian carcinoma (OVCA) LYP Bestatin Growth inhibition
This project was supported by the Natural Science Foundation of China (30973550) and Shandong Provincial Foundation for Natural Science (Y2008C07). This work was also supported by Department of Pathology and Laboratory Medicine, University of Calgary and Calgary Laboratory Services, Canada.
- 5.Reinhold D, Bank U, Täger M, Ansorge S, Wrenger S, Thielitz A, Lendeckel U, Faust J, Neubert K, Brocke S (2008) DP IV/CD26, APN/CD13 and related enzymes as regulators of T cell immunity: implications for experimental encephalomyelitis. Front Biosci 13:2356–2363. doi: org/10.2741/2849 PubMedCrossRefGoogle Scholar
- 9.Wang C, Fan G, Lin M, Chen Y, Zhao W, Wu Y (2007) Development of a liquid chromatography/tandem mass spectrometry assay for the determination of bestatin in rat plasma and its application to a pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci 850:101–108. doi: 10.1016/j.jchromb.2006.11.015 PubMedCrossRefGoogle Scholar
- 14.Chandrappa S, Benaka Prasad SB, Vinaya K, Ananda Kumar CS, Thimmegowda NR, Rangappa KS (2008) Synthesis and in vitro antiproliferative activity against human cancer cell lines of novel 5-(4-methyl-benzylidene)-thiazolidine-2, 4-diones. Invest New Drugs 26:437–444. doi: 10.1007/s10637-008-9130-7 PubMedCrossRefGoogle Scholar
- 15.Shahabuddin MS, Nambiar M, Choudhary B, Advirao GM, Raghavan SC (2010) A novel DNA intercalator, butylamino-pyrimido[4′, 5′:4, 5]selenolo(2, 3-b)quinoline, induces cell cycle arrest and apoptosis in leukemic cells. Invest New Drugs 28:35–48. doi: 10.1007/s10637-008-9212-6 PubMedCrossRefGoogle Scholar
- 19.Liu J, Li X, Cheng YN, Cui SX, Chen MH, Xu WF, Tian ZG, Makuuchi M, Tang W, Qu XJ (2007) Inhibition of human gastric carcinoma cell growth by treatment of N(3)-o-toluyl-fluorouracil as a precursor of 5-fluorouracil. Eur J Pharmacol 574:1–7. doi: 10.1016/j.ejphar.2007.06.064 PubMedCrossRefGoogle Scholar
- 28.Ito K, Nakajima Y, Onohara Y, Takeo M, Nakashima K, Matsubara F, Ito T, Yoshimoto T (2006) Crystal structure of aminopeptidase N (proteobacteria alanyl aminopeptidase) from Escherichia coli and conformational change of methionine 260 involved in substrate recognition. J Biol Chem 281:33664–33676. doi: 10.1074/jbc.M605203200 PubMedCrossRefGoogle Scholar