title: ROS-dependent antiproliferative effect of brassinin derivative homobrassinin in human colorectal cancer Caco2 cells.
abstract: This study was designed to examine the in vitro antiproliferative effect of brassinin and its derivatives on human cancer cell lines. Among seven tested compounds, homobrassinin (K1; N-[2-(indol-3-yl)ethyl]-S-methyldithiocarbamate) exhibited the most potent activity with IC50 = 8.0 μM in human colorectal Caco2 cells and was selected for further studies. The flow cytometric analysis revealed a K1-induced increase in the G2/M phase associated with dysregulation of α-tubulin, α1-tubulin and β5-tubulin expression. These findings suggest that the inhibitory effect of K1 can be mediated via inhibition of microtubule formation. Furthermore, simultaneously with G2/M arrest, K1 also increased population of cells with sub-G1 DNA content which is considered to be a marker of apoptotic cell death. Apoptosis was also confirmed by annexin V/PI double staining, DNA fragmentation assay and chromatin condensation assay. The apoptosis was associated with the loss of mitochondrial membrane potential (MMP), caspase-3 activation as well as intracellular reactive oxygen species (ROS) production. Moreover, the antioxidant Trolox blocked ROS production, changes in MMP and decreased K1 cytotoxicity, which confirmed the important role of ROS in cell apoptosis. Taken together, our data demonstrate that K1 induces ROS-dependent apoptosis in Caco2 cells and provide the rationale for further in vivo anticancer investigation.
authors: Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: Martin
authors: M
authors: Kello
authors: Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: David
authors: D
authors: Drutovic
authors: Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: Martina
authors: M
authors: Chripkova
authors: Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: Martina
authors: M
authors: Pilatova
authors: Department of Organic Chemistry, Institute of Chemical Sciences, Faculty of Science, Pavol Jozef Safarik University, 040 80 Kosice, Slovak Republic.
authors: Mariana
authors: M
authors: Budovska
authors: Department of Experimental Medicine, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: Lucia
authors: L
authors: Kulikova
authors: Department of Gynaecology and Obstetrics, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic.
authors: Peter
authors: P
authors: Urdzik
authors: Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, 040 11 Kosice, Slovak Republic. jan.mojzis@upjs.sk.
authors: Jan
authors: J
authors: Mojzis
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D018938:Caco-2 Cells; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000737:chemistry / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013859:Thiocarbamates / Q000737:chemistry / Q000494:pharmacology*; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D000094162:Phytoalexins
chemical_list: D000970:Antineoplastic Agents; D053148:Caspase 3; D007211:Indoles; D017382:Reactive Oxygen Species; D013859:Thiocarbamates; D014404:Tubulin; C089020:brassinin; D000094162:Phytoalexins