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Novel Benzo[B]Furans with Anti-Microtubule Activity Upregulate Expression of Apoptotic Genes and Arrest Leukemia Cells in G2/M Phase

[ Vol. 19 , Issue. 3 ]

Author(s):

Karolina Królewska-Goliñska, Marcin J. Cieślak*, Milena Sobczak, Rafał Dolot, Ewa Radzikowska-Cieciura, Mariola Napiórkowska, Iwona Wybrañska and Barbara Nawrot   Pages 375 - 388 ( 14 )

Abstract:


Background: Novel derivatives of benzo[b]furan were found to be highly toxic towards human chronic myelogenous (K562), acute myelogenous (HL-60) and acute lymphoblastic (MOLT-4) leukemia cells.

Objective: Characterization of the biological activity of novel benzofurans (influence on apoptosis, mitogen activated protein kinases and on the cell cycle). Identification of cellular protein(s) targeted by test benzofurans and defining the mechanism of action.

Method: chemical synthesis, fluorescence assays, flow cytometry, gene expression by DNA microarray and real time RT-PCR, western blotting, cytotoxicity assays, pull-down assay, mass spectroscopy, in vitro polymerization of tubulin, molecular docking.

Results: 1,1'-[3-(bromomethyl)-5,6- dimethoxy-1-benzofuran-2,7-diyl]diethanone (1) and methyl 4-bromo-6-(dibromoacetyl)-5-hydroxy-2-methyl-1-benzofuran-3-carboxylate (2) induced apoptosis in K562 and MOLT-4 cells. The profiling of gene expression revealed that 1 and 2 increased the expression of proapoptotic genes involved in both receptor (TNFRSF 10A, TNFRSF 10B, CASP8) and mitochondrial (BAX, BID, NOXA, APAF1) pathways of apoptosis. Test benzo[b]furans activated c-Jun N-terminal kinase (JNK) and p38 kinase in K562 cells. Tubulin was identified as a protein target for benzo[b]furans in pull-down experiments with biotinylated 2. Test benzo[b]furans inhibited polymerization of tubulin monomers in vitro, decreased the level of cellular microtubules and arrested cells in a G2/M phase. Molecular docking suggests that benzo[b]furans 1 and 2 bind to tubulin via colchicine binding pocket and the complex is stabilized mainly by hydrophobic interactions.

Conclusion: Novel benzo[b]furans with anti-microtubule activity were identified. They induce apoptosis in cancer cells and cause G2/M cell cycle arrest. Biological activity of 1 and 2 makes them potential lead compounds for development as anticancer drugs.

Keywords:

Apoptosis, benzofurans, microtubules, antineoplastic agents, leukemia, molecular docking, novel benzo[b]furans.

Affiliation:

Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz, Department of Medical Chemistry, Medical University of Warsaw, 3 Oczki Str., 02-007 Warsaw, Department of Genetic Diagnostics and Nutrigenomics, Chair of Clinical Biochemistry, The Jagiellonian University, Medical College, 15 Kopernika Str., 31-501 Krakow, Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Department of Bioorganic Chemistry, 112 Sienkiewicza Str., 90-363 Lodz



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