title: Effect of diterpenoid kaurenoic acid on genotoxicity and cell cycle progression in gastric cancer cell lines.
abstract: The goal of our study was to evaluate the effect of kaurenoic acid, obtained from copaiba oil resin, in gastric cancer (GC) and a normal mucosa of stomach (MNP01) cell lines. The compound was tested at concentrations of 2.5, 5, 10, 30 and 60μg/mL. Comet and micronucleus assays were used to access its potential genotoxicity in vitro. Moreover, we evaluated the effect of kaurenoic acid in cell cycle progression and in the transcription of genes involved in the control of the cell cycle: MYC, CCND1, BCL2, CASP3, ATM, CHK2 and TP53. Kaurenoic acid induced an increase on cell DNA damage or micronucleus frequencies on GC cell lines in a dose-dependent manner. The GC and MNP01 cell lines entering DNA synthesis and mitosis decreased significantly with kaurenoic acid treatment, and had an increased growth phase compared with non-treated cells. The treatment induced apoptosis (or necrosis) even at a concentration of 2.5μg/mL in relation to non-treated cells. GC cell lines presented reduced MYC, CCND1, BCL2 and CASP3 transcription while ATM, CHK2 and TP53 increased in transcription in relation to non-treated cells, especially at a concentration above 10μg/mL. The gene transcription in the MNP01 (non-treated non-cancer cell line) was designated as a calibrator for all the GC cell lines. In conclusion, our results showed that kaurenoic acid obtained from Copaifera induces DNA damage and increases the micronuclei frequency in a dose-dependent manner in GC cells, with a significant genotoxicity observed above the concentration of 5μg/mL. Moreover, this compound seems to be able to induce cell cycle arrest and apoptosis in GC cells.
authors.affiliation: Human Cytogenetic Laboratory, Biological Science Institute, Federal University of Pará (UFPA), Belém, Pará, Brazil.
authors.forename: Plínio Cerqueira Dos Santos
authors.initials: PCDS
authors.lastname: Cardoso
authors.affiliation: Federal Institute of Education, Science and Technology of Pará (IFPA), Av. Almirante Barroso, 1155 (Marco), CEP 66093-020, Belém, Pará, Brazil. Electronic address: carlos.rocha@ifpa.edu.br.
authors.forename: Carlos Alberto Machado da
authors.initials: CAMD
authors.lastname: Rocha
authors.affiliation: Department of Morphology and Genetics, Federal University of São Paulo (UNIFESP), São Paulo, São Paulo, Brazil.
authors.forename: Mariana Ferreira
authors.initials: MF
authors.lastname: Leal
authors.affiliation: Human Cytogenetic Laboratory, Biological Science Institute, Federal University of Pará (UFPA), Belém, Pará, Brazil.
authors.forename: Marcelo de Oliveira
authors.initials: MO
authors.lastname: Bahia
authors.affiliation: Human Cytogenetic Laboratory, Biological Science Institute, Federal University of Pará (UFPA), Belém, Pará, Brazil.
authors.forename: Diego Di Felipe Ávila
authors.initials: DDFÁ
authors.lastname: Alcântara
authors.affiliation: Laboratory of Genetics and Molecular Biology, University of Franca (UNIFRAN), Franca, São Paulo, Brazil.
authors.forename: Raquel Alves Dos
authors.initials: RAD
authors.lastname: Santos
authors.affiliation: Laboratory of Genetics and Molecular Biology, University of Franca (UNIFRAN), Franca, São Paulo, Brazil.
authors.forename: Natália Dos Santos
authors.initials: NDS
authors.lastname: Gonçalves
authors.affiliation: Laboratory of Biotransformation, University of Franca (UNIFRAN), Franca, São Paulo, Brazil.
authors.forename: Sérgio Ricardo
authors.initials: SR
authors.lastname: Ambrósio
authors.affiliation: Department of Physiology and Pharmacology, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Bruno Coêlho
authors.initials: BC
authors.lastname: Cavalcanti
authors.affiliation: Human Cytogenetic Laboratory, Biological Science Institute, Federal University of Pará (UFPA), Belém, Pará, Brazil; Laboratory of Genetics of Hemoglobinopathies and Hematologic Diseases, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Caroline Aquino
authors.initials: CA
authors.lastname: Moreira-Nunes
authors.affiliation: Department of Physiology and Pharmacology, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil.
authors.forename: Claudia do Ó
authors.initials: CDÓ
authors.lastname: Pessoa
authors.affiliation: Human Cytogenetic Laboratory, Biological Science Institute, Federal University of Pará (UFPA), Belém, Pará, Brazil; Hospital Ophir Loyola (HOL), Belém, Pará, Brazil. Electronic address: rommel@ufpa.br.
authors.forename: Rommel Mário Rodríguez
authors.initials: RMR
authors.lastname: Burbano
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects*; D002460:Cell Line; D004224:Diterpenes / Q000737:chemistry / Q000494:pharmacology*; D005753:Gastric Mucosa / Q000166:cytology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D015394:Molecular Structure; D009152:Mutagenicity Tests; D013274:Stomach Neoplasms
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D004224:Diterpenes; C051436:kaurenoic acid
keywords: Cell cycle progression; Comet assay; Gastric cancer cell lines; Gene transcriptions; Kaurenoic acid; Micronuclei