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Hohla, Florian; Buchholz, Stefan; Schally, Andrew V; Seitz, Stefan; Rick, Ferenc G.; Szalontay, Luca; Varga, Jozsef L.; Zarandi, Marta; Halmos, Gabor; Vidaurre, Irving; Krishan, Awtar; Kurtoglu, Metin; Chandna, Sudhir; Aigner, Elmar; Datz, Christian
Cell cycle (Georgetown, Tex.), 10/1/2009, 2009/10/01, 2009-Oct-01, 2009-10-00, 20091001, Letnik: 8, Številka: 19Journal Article
We investigated the mechanisms of inhibitory effect of growth hormone-releasing hormone (GHRH) antagonist JMR-132 on the growth of HT29, HCT-116 and HCT-15 human colon cancer cells in vitro and in vivo. High-affinity binding sites for GHRH and mRNA for GHRH and splice variant-1 (SV1) of the GHRH receptor were found in all three cell lines tested. Proliferation of HT-29, HCT-116 and HCT-15 cells was significantly inhibited in vitro by JMR-132. Time course studies revealed that the treatment of human HCT-116 colon cancer cells with 10μM GHRH antagonist JMR-132 causes a significant DNA damage as shown by an increase in olive tail moment (OTM) and loss of inner mitochondrial membrane potential (∆Ψm). Western blotting demonstrated a time-dependent increase in protein levels of phospho-p53 (Ser46), Bax, cleaved caspase-9, -3, cleavage of poly(ADP-ribose)polymerase (PARP) and a decrease in Bcl-2 levels. An augmentation in cell cycle checkpoint protein p21Waf1/Cip1 was accompanied by a cell cycle arrest in S-phase. DNA fragmentation visualized by the comet assay and the number of apoptotic cells increased time dependently as determined by flow cytometric annexinV and PI staining assays. In vivo, JMR-132 decreased the volume of HT-29, HCT-116 and HCT-15 tumors xenografted into athymic mice up to 75% (p
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in: SICRIS
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