title: Morphological and biochemical alterations activated by antitumor clerodane diterpenes.
abstract: Casearia sylvestris Swartz (Salicaceae) is a plant commonly widespread in the Americas. It has oxygenated tricyclic bioactive clerodane diterpenes with antimicrobial, antiulcer, larvicidal, chemopreventive, anti-inflammatory, antioxidant and antiproliferative properties. Due to this requirement for the developing of new anticancer drugs, it was initially evaluated the cytotoxic activity of a fraction with Casearins (FC) and its clerodane diterpenes Casearin B (Cas B), D (Cas D), X (Cas X) and Caseargrewiin F (Cas F) isolated from C.sylvestris leaves against 7 tumor cell lines, Sarcoma 180 cells (S180) and on normal peripheral blood mononuclear cells (PBMC). All substances tested showed cytotoxic potential. Cas F and X were the most active compounds. Cell death analyzes with Cas F (0.5 and 1μM) and Cas X (0.7 and 1.5μM) using the HL-60 leukemia line as experimental model showed DNA synthesis and membrane integrity reduction, DNA fragmentation and mitochondrial depolarization, specially after 24h exposure, cell cycle arrest in G0/G1 phase caused by Cas X, activation of the initiator -8/-9 and effector -3/-7 caspases and phosphatidylserine externalization, all biochemical features of apoptosis corroborated by chromatinic condensation, karyorrhexis, cytoplasmic vacuolation and rarefaction and cellular shrinkage, morphological findings specially observed after 12 and 24h of incubation. Therefore, Cas X and F were the most functional molecules with more pronounced lethal and discriminating effects on tumor cells and antiproliferative action predominantly mediated by apoptosis, highlighting clerodane dipertenes as promising lead antineoplastic compounds.
authors.affiliation: Department of Biophysics and Physiology, Campus Ministro Petrônio Portella, Federal University of Piauí, Teresina, Brazil; Postgraduate Program in Pharmaceutical Sciences, Federal University of Piauí, Teresina, Brazil. Electronic address: pmpf@ufpi.edu.br.
authors.forename: Paulo Michel Pinheiro
authors.initials: PM
authors.lastname: Ferreira
authors.affiliation: Department of Physiology and Pharmacology, Federal University of Pernambuco, Recife, Brazil.
authors.forename: Gardenia Carmen Gadelha
authors.initials: GC
authors.lastname: Militão
authors.affiliation: Department of Physiology and Pharmacology, Faculty of Medicine, Federal University of Ceará, Fortaleza, Brazil.
authors.forename: Daisy Jereissati Barbosa
authors.initials: DJ
authors.lastname: Lima
authors.affiliation: Department of Biological Sciences, Campus Senador Helvídio Nunes de Barros, Federal University of Piauí, Picos, Brazil.
authors.forename: Nagilla Daniela de Jesus
authors.initials: ND
authors.lastname: Costa
authors.affiliation: Postgraduate Program in Pharmaceutical Sciences, Federal University of Piauí, Teresina, Brazil.
authors.forename: Kátia da Conceição
authors.initials: Kda C
authors.lastname: Machado
authors.affiliation: Faculty of Pharmaceutical Sciences, State University of São Paulo Júlio de Mesquita Filho, Araraquara, Brazil.
authors.forename: André Gonzaga Dos
authors.initials: AG
authors.lastname: Santos
authors.affiliation: Chemistry Institute, State University of São Paulo Júlio de Mesquita Filho, Araraquara, Brazil.
authors.forename: Alberto José
authors.initials: AJ
authors.lastname: Cavalheiro
authors.affiliation: Chemistry Institute, State University of São Paulo Júlio de Mesquita Filho, Araraquara, Brazil.
authors.forename: Vanderlan da Silva
authors.initials: Vda S
authors.lastname: Bolzani
authors.affiliation: Chemistry Institute, State University of São Paulo Júlio de Mesquita Filho, Araraquara, Brazil.
authors.forename: Dulce Helena Siqueira
authors.initials: DH
authors.lastname: Silva
authors.affiliation: Department of Physiology and Pharmacology, Faculty of Medicine, Federal University of Ceará, Fortaleza, Brazil.
authors.forename: Cláudia
authors.initials: C
authors.lastname: Pessoa
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D037821:Casearia / Q000737:chemistry; D020169:Caspases / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D053938:DNA Fragmentation; D004224:Diterpenes / Q000494:pharmacology; D045785:Diterpenes, Clerodane / Q000737:chemistry / Q000494:pharmacology*; D005260:Female; D018922:HL-60 Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D010718:Phosphatidylserines / Q000378:metabolism; D018515:Plant Leaves / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism; D012510:Sarcoma 180
chemical_list: C548433:1-(acetyloxy)-3,5,6,6a,7,8,9,10-octahydro-10-hydroxy-7,8-dimethyl-7-(3-methylpenta-2,4-dien-1-yl)naphtho(1,8a-c)furan-3,5-diyl dibutanoate; D000972:Antineoplastic Agents, Phytogenic; D004224:Diterpenes; D045785:Diterpenes, Clerodane; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species; C546626:caseargrewiin F; C582885:casearin B; C000614112:casearin D; D020169:Caspases
keywords: Antiproliferative action; Apoptosis; Casearia sylvestris; Cytotoxicity; Human cells; Murine cells