Abstract
The aim of the present study was to assess the cytotoxicity of manumycin, a specific inhibitor of farnesyl:protein transferase, as well as its effects on protein isoprenylation and kinase-dependent signal transduction in COLO320-DM human colon adenocarcinoma which harbours a wild-type K- ras gene. Immunoblot analysis of isolated cell membranes and total cellular lysates of COLO320-DM cells demonstrated that manumycin dose-dependently reduced p21 ras farnesylation with a 50% inhibitory concentration (IC50) of 2.51 ± 0.11 μM and 2.68 ± 0.20 μM, respectively, while the geranylgeranylation of p21 rhoA and p21 rap1 was not affected. Manumycin dose-dependently inhibited (IC50= 2.40 ± 0.67 μM) the phosphorylation of the mitogen-activated protein kinase/extracellular-regulated kinase 2 (p42MAPK/ERK2), the main cytoplasmic effector of p21 ras, as well as COLO320-DM cell growth (IC50= 3.58 ± 0.27 μM) without affecting the biosynthesis of cholesterol. Mevalonic acid (MVA, 100 μM), a substrate of the isoprenoid synthesis, was unable to protect COLO320-DM cells from manumycin cytotoxicity. Finally, manumycin 1–25 μM for 24–72 h induced oligonucleosomal fragmentation in a dose- and time-dependent manner and MVA did not protect COLO320-DM cells from undergoing DNA cleavage. The present findings indicate that the inhibition of p21 ras processing and signal transduction by manumycin is associated with marked inhibition of cell proliferation and apoptosis in colon cancer cells and the effect on cell growth does not require the presence of a mutated ras gene for maximal expression of chemotherapeutic activity. © 2000 Cancer Research Campaign
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References
Bos JL (1989) Ras oncogenes in human cancer: a review. Cancer Res 49: 4682–4689
Corsini A, Maggi FM and Catapano AL (1995) Pharmacology of competitive inhibitors of HMG-CoA reductase. Pharmacol Res 31: 9–27
Cox AD and Der CJ (1997) Farnesyltransferase inhibitors and cancer treatment: targeting simply Ras?. Biochim Biophys Acta 1333: F51–F71
Danesi R, Figg WD, Reed E and Myers CE (1995) Paclitaxel (Taxol®) inhibits protein isoprenylation and induces apoptosis in PC-3 human prostate cancer cells. Mol Pharmacol 47: 1106–1111
Danesi R, Nardini D, Basolo F, Del Tacca M, Samid D and Myers CE (1996) Phenylacetate inhibits protein isoprenylation and growth of the androgen-independent LNCaP prostate cancer cells transfected with the T24 Ha-ras oncogene. Mol Pharmacol 49: 972–979
Giancotti FG and Mainiero F (1994) Integrin-mediated adhesion and signaling in tumorigenesis. Biochim Biophys Acta 1198: 47–64
Giannakouros T and Magee AI (1993) Protein prenylation and associated modifications. In: Lipid Modifications of Proteins, Schlesinger MJ (ed), pp. 136–162, CRC Press: Boca Raton, FL
Gibbs JB and Oliff A (1997) The potential of farnesyltransferase inhibitors as cancer chemotherapeutics. Annu Rev Pharmacol Toxicol 37: 143–146
Gibbs JB, Oliff A and Kohl NE (1994) Farnesyltransferase inhibitors: ras research yields a potential cancer therapeutics. Cell 77: 175–178
Hara M and Han M (1995) Ras farnesyltransferase inhibitors suppress the phenotype resulting from an activated ras mutation in Caenorhabditis elegans. Proc Natl Acad Sci USA 92: 3333–3337
Hara M, Kazuhito A, Akinaga S, Okabe M, Nakano H, Gomez R, Wood D, Uh M and Tamanoi F (1993) Identification of ras farnesyltransferase inhibitors by microbial screening. Proc Natl Acad Sci USA 90: 2281–2285
Ikeda M, Koyama S, Okazaki M, Dohi K and Kikuchi A (1995) Rap1 p21 regulates the interaction of ras p21 with RGL, a new effector protein of ras p21. FEBS Lett 375: 37–40
Kawabata H, Anzai N, Yoshida Y and Okuma M (1994) A new method for quantitative estimation of the degree of DNA fragmentation utilizing agarose gel electrophoresis. Int J Hematol 59: 311–316
Larcher F, Robles AI, Duran H, Murillas R, Quintanilla M, Cano A, Conti CJ and Jorcano JL (1996) Up-regulation of vascular endothelial growth factor/vascular permeability factor in mouse skin carcinogenesis correlates with malignant progression state and activated H- ras expression levels. Cancer Res 56: 5391–5396
Levitzky A (1996) Targeting signal transduction for disease therapy. Curr Opin Cell Biol 8: 239–244
Lowry OH, Rosebrough NJ, Farr AL and Randall RJ (1951) Protein measurement with folin phenol reagent. J Biol Chem 193: 265–267
Lowy DR and Willumsen BM (1993) Function and regulation of ras. Annu Rev Biochem 62: 851–891
Maltese WA (1990) Post-translational modification of proteins by isoprenoids in mammalian cells. FASEB J 4: 3319–3328
Marchetti A, Buttitta F, Pellegrini S, Chella A, Bertacca G, Filardo A, Tognoni V, Ferreli F, Signorini E, Angeletti CA and Bevilacqua G (1996) Bronchioloalveolar lung carcinomas: K- ras mutations constant events in the mucinous subtype. J Pathol 179: 254–259
Marshall CJ (1993) Protein prenylation: a mediator of protein–protein interactions. Science 259: 1865–1866
Marshall CJ (1996) Ras effectors. Curr Opin Cell Biol 8: 197–204
Marshall MS (1995) Ras target proteins in eukaryotic cells. FASEB J 9: 1311–1318
Nagase T, Kawata S, Tamura S, Matsuda Y, Inui Y, Yamasaky E, Ishiguro H, Ito T and Matsuzawa Y (1996) Inhibition of cell growth of human hepatoma cell line (Hep G2) by a farnesyl protein transferase inhibitor: a preferential suppression of ras farnesylation. Int J Cancer 65: 620–626
Nagasu T, Yoshimatsu K, Rowell C, Lewis MD and Garcia AM (1995) Inhibition of human tumor xenograft growth by treatment with farnesyl transferase inhibitor B956. Cancer Res 55: 5310–5314
Nassar N, Horn G, Hermann C, Scherer A, McCormick F and Wittinghofer A (1995) The 2.2 Å crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with Rap1A and a GTP analogue. Nature 375: 554–560
Perez-Sala D and Mollinedo F (1994) Inhibition of isoprenoid biosynthesis induces apoptosis in human promyelocytic HL-60 cells. Biochem Biophys Res Commun 199: 1209–1215
Phan T, McLeod JG, Pollard JD, Peiris O, Rohan A and Halpern J-P (1995) Peripheral neuropathy associated with simvastatin. J Neurol Neurosurg Psychiatry 58: 625–628
Rodenhuis S, Boerrigter L, Top B, Slebos RJC, Mooi WJ, van’t Veer L and van Zandwijk N (1997) Mutational activation of the K- ras oncogene and the effect of chemotherapy in advanced adenocarcinoma of the lung: a prospective study. J Clin Oncol 15: 285–291
Tamanoi F (1993) Inhibitors of Ras farnesyltransferases. Trends Biol Sci 18: 349–353
Veerkamp JH, Smith JWA, Benders AAGM and Oosterhof A (1996) Effects of HMG-CoA reductase inhibitors on growth and differentiation of cultured rat skeletal muscle cells. Biochim Biophys Acta 1315: 217–223
Wittinghofer A and Herrman C (1995) Ras -effector interactions, the problem of specificity. FEBS Lett 369: 52–56
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Paolo, A., Danesi, R., Nardini, D. et al. Manumycin inhibits ras signal transduction pathway and induces apoptosis in COLO320-DM human colon tumourcells. Br J Cancer 82, 905–912 (2000). https://doi.org/10.1054/bjoc.1999.1018
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DOI: https://doi.org/10.1054/bjoc.1999.1018
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