title: Synthesis, characterization and systematic comparison of FITC-labelled GnRH-I, -II and -III analogues on various tumour cells.
abstract: Targeted tumour therapy is the focus of recent cancer research. Gonadotropin-releasing hormone (GnRH) analogues are able to deliver anticancer agents selectively into tumour cells, which highly express GnRH receptors. However, the effectiveness of different analogues as targeting moiety in drug delivery systems is rarely compared, and the investigated types of cancer are also limited. Therefore, we prepared selectively labelled, fluorescent derivatives of GnRH-I, -II and -III analogues, which were successfully used for drug targeting. In this manuscript, we investigated these analogues' solubility, stability and passive membrane permeability and compared their cellular uptake by various cancer cells. We found that these labelled GnRH conjugates provide great detectability, without undesired cytotoxicity and passive membrane permeability. The introduced experiments with these conjugates proved their reliable tracking, quantification and comparison. Cellular uptake efficiency was studied on human breast, colon, pancreas and prostate cancer cells (MCF-7, HT-29, BxPC-3, LNCaP) and on dog kidney cells (Madin-Darby canine kidney). Each of the three conjugates was taken up by GnRH-I receptor-expressing cells, but the different cells preferred different analogues. Furthermore, we demonstrated for the first time the high cell surface expression of GnRH-I receptors and the effective cellular uptake of GnRH analogues on human pharynx tumour (Detroit-562) cells. In summary, our presented results detail that the introduced conjugates could be innovative tools for the examination of the GnRH-based drug delivery systems on various cells and offer novel information about these peptides. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.|Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: József
authors.initials: J
authors.lastname: Murányi
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: Pál
authors.initials: P
authors.lastname: Gyulavári
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: Attila
authors.initials: A
authors.lastname: Varga
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: Györgyi
authors.initials: G
authors.lastname: Bökönyi
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: Henriette
authors.initials: H
authors.lastname: Tanai
authors.affiliation: Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: Tibor
authors.initials: T
authors.lastname: Vántus
authors.affiliation: 1st Department of Pediatrics, Semmelweis University, Bókay János St. 53-54, H1083, Budapest, Hungary.
authors.forename: Domonkos
authors.initials: D
authors.lastname: Pap
authors.affiliation: Department of Pharmaceutics, Semmelweis University, Hőgyes Endre St. 7, H1092, Budapest, Hungary.
authors.forename: Krisztina
authors.initials: K
authors.lastname: Ludányi
authors.affiliation: MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Eötvös L. University, Pázmány Péter sétány 1/A, H1518, Budapest, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Mező
authors.affiliation: MTA-SE Pathobiochemistry Research Group, Tűzoltó St. 37-47, H1094, Budapest, Hungary.|Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Tűzoltó St. 37-47, H1094, Budapest, Hungary.
authors.forename: György
authors.initials: G
authors.lastname: Kéri
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D002470:Cell Survival / Q000187:drug effects; D004285:Dogs; D004337:Drug Carriers; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D015870:Gene Expression; D007987:Gonadotropin-Releasing Hormone / Q000138:chemical synthesis* / Q000235:genetics / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D007700:Kinetics; D061986:MCF-7 Cells; D061985:Madin Darby Canine Kidney Cells; D008297:Male; D009928:Organ Specificity; D020033:Protein Isoforms / Q000138:chemical synthesis / Q000235:genetics / Q000378:metabolism; D011966:Receptors, LHRH / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D013194:Staining and Labeling / Q000379:methods
chemical_list: D004337:Drug Carriers; D005456:Fluorescent Dyes; D020033:Protein Isoforms; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone; D016650:Fluorescein-5-isothiocyanate
keywords: Detroit-562; FITC; GnRH; LHRH; conjugate; pharynx tumour; targeted therapy