Melatonin and the MT1 and MT2 receptors: effects on apoptosis, cell proliferation and migratory potential of the ovarian carcinoma cells (SKOV-3 cell line)

• Grant number: 19/00906-6
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2019
• End date: September 30, 2021
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Luiz Gustavo de Almeida Chuffa
• Grantee: Luiz Gustavo de Almeida Chuffa
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated researchers: Graziela Gorete Romagnoli ; Lucilene Delazari dos Santos ; Ramon Kaneno ; Robson Francisco Carvalho

Abstract

Ovarian cancer (OC) is the most lethal gynecological malignancy and the fifth leading cause of cancer death, mainly due to the resistance to treatments associated with poor prognosis. Melatonin (Mel), an indoleamine secreted by the pineal gland in the circadian cycle dark phase, has shown a number of antitumor properties both in vitro and in vivo. Evidences suggest that Mel exhibit an oncostatic effect on several cancer types (e.g. OC), and this effect seems to be mediated by its membrane receptors, termed MT1 and MT2. This seems to have a special relationship with mitochondria, which is responsible for its synthesis, energy metabolism, cell death and differentiation. The aim of the present study is to evaluate the influence of melatonin therapy on the endogenous mitochondrial signaling pathway and apoptosis of human ovarian carcinoma cells (SKOV-3), and also to verify whether the biological effects are dependent on the activation of MT1/2 receptors through the RNAs (siRNAs) knockdown. SKOV-3 cells will be silenced for MT1/2 receptors followed by the treatment with Mel (800 ¼M, 1600 ¼M and 2400 ¼M) or not. Control groups will be set for each treatment. Cell adhesion proteins, cell migration and invasion, cytotoxicity, cell cycle arresting as well as apoptosis will be carried out by MTT assay, transwell inserts, multiplex assay, western blotting and flow cytometry. For apoptosis, Annexin-V will be assayed in association with proteins such as Bcl-2, BAX, cytochrome c, and cleaved caspase-3. In addition, mitochondria will be isolated from SKOV-3 cells to effectively access the melatonin levels, energy metabolism, and expression of proteins involved with endogenous melatonin synthesis (AA-NAT and ASMT). By using proteomic analysis through electrospray (LC/MS-MS), global proteins and peptides will be investigated in these cells. These findings may contribute to the current discussion on the role of melatonin in the altered metabolism of neoplastic OC cells. (AU)

Scientific publications

(13)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LUPI, LUIZ ANTONIO; DELELLA, FLAVIA KARINA; CUCIELO, MAIRA SMANIOTTO; ROMAGNOLI, GRAZIELA GORETE; KANENO, RAMON; NUNES, ISEU DA SILVA; DOMENICONI, RAQUEL FANTIN; MARTINEZ, MARCELO; MARTINEZ, FRANCISCO EDUARDO; FAVARO, WAGNER JOSA; et al. P-MAPA and Interleukin-12 Reduce Cell Migration/Invasion and Attenuate the Toll-Like Receptor-Mediated Inflammatory Response in Ovarian Cancer SKOV-3 Cells: A Preliminary Study. Molecules, v. 25, n. 1, JAN 2020. (16/03993-9, 19/00906-6)

DE ALMEIDA CHUFFA, LUIZ GUSTAVO; LUPI, LUIZ ANTONIO; CUCIELO, MAIRA SMANIOTTO; SILVEIRA, HENRIQUE SPAULONCI; REITER, RUSSEL J.; FERREIRA SEIVA, FABIO RODRIGUES. Melatonin Promotes Uterine and Placental Health: Potential Molecular Mechanisms. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 21, n. 1, JAN 1 2020. (19/00906-6)

LUPI, LUIZ ANTONIO; CUCIELO, MAIRA SMANIOTTO; SILVEIRA, HENRIQUE SPAULONCI; GAIOTTE, LETICIA BARBOSA; CESARIO, ROBERTA CARVALHO; FERREIRA SEIVA, FABIO RODRIGUES; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. The role of Toll-like receptor 4 signaling pathway in ovarian, cervical, and endometrial cancers. Life Sciences, v. 247, APR 15 2020. (19/00906-6)

DE MORAIS OLIVEIRA, DIEGO AUGUSTO; LUPI, LUIZ ANTONIO; SILVEIRA, HENRIQUE SPAULONCI; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. Protein restriction during puberty alters nutritional parameters and affects ovarian and uterine histomorphometry in adulthood in rats. International Journal of Experimental Pathology, MAR 2021. (19/00906-6, 18/09243-7)

SILVEIRA, HENRIQUE SPAULONCI; LUPI, LUIZ ANTONIO; ROMAGNOLI, GRAZIELA GORETE; KANENO, RAMON; NUNES, ISEU DA SILVA; FAVARO, WAGNER JOSE; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. P-MAPA activates TLR2 and TLR4 signaling while its combination with IL-12 stimulates CD4+and CD8+effector T cells in ovarian cancer. Life Sciences, v. 254, AUG 1 2020. (17/03441-9, 16/03993-9, 19/00906-6)

DE MORAIS OLIVEIRA, DIEGO AUGUSTO; LUPI, LUIZ ANTONIO; SILVEIRA, HENRIQUE SPAULONCI; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. Protein restriction during puberty alters nutritional parameters and affects ovarian and uterine histomorphometry in adulthood in rats. International Journal of Experimental Pathology, v. 102, n. 2, p. 93-104, APR 2021. (18/09243-7, 19/00906-6)

CHUFFA, L. G.; CUCIELO, M.; CARVALHO, R. F.; ROMAGNOLI, G. G.; KANENO, R.; SEIVA, F.. Melatonin reverses the Warburg-dependent effect in ovarian cancer cell by binding to the MT1 and MT2 receptors. Annals of Oncology, v. 31, p. 2-pg., 2020-09-01. (19/00906-6, 18/15797-5)

CHUFFA, LUIZ GUSTAVO DE ALMEIDA; CARVALHO, ROBSON FRANCISCO; JUSTULIN, LUIS ANTONIO; CURY, SARAH SANTILONI; SEIVA, FABIO RODRIGUES FERREIRA; JARDIM-PERASSI, BRUNA VICTORASSO; ZUCCARI, DEBORA APARECIDA PIRES DE CAMPOS; REITER, RUSSEL J.. A meta-analysis of microRNA networks regulated by melatonin in cancer: Portrait of potential candidates for breast cancer treatment. Journal of Pineal Research, v. 69, n. 4, NOV 2020. (19/00906-6)

DE ALMEIDA CHUFFA, LUIZ GUSTAVO; FERREIRA SEIVA, FABIO RODRIGUES; NOVAIS, ADRIANA ALONSO; SIMAO, VINICIUS AUGUSTO; MARTIN GIMENEZ, VIRNA MARGARITA; MANUCHA, WALTER; PIRES DE CAMPOS ZUCCARI, DEBORA APARECIDA; REITER, RUSSEL J.. Melatonin-Loaded Nanocarriers: New Horizons for Therapeutic Applications. Molecules, v. 26, n. 12, JUN 2021. (19/00906-6)

LUPI JUNIOR, LUIZ ANTONIO; CUCIELO, MAIRA SMANIOTTO; DOMENICONI, RAQUEL FANTIN; DOS SANTOS, LUCILENE DELAZARI; SILVEIRA, HENRIQUE SPAULONCI; NUNES, ISEU DA SILVA; MARTINEZ, MARCELO; MARTINEZ, FRANCISCO EDUARDO; FAVARO, WAGNER JOSE; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. P-MAPA and IL-12 Differentially Regulate Proteins Associated with Ovarian Cancer Progression: A Proteomic Study. ACS OMEGA, v. 4, n. 26, p. 21761-21777, DEC 24 2019. (16/03993-9, 19/00906-6)

CESARIO, ROBERTA CARVALHO; GAIOTTE, LETICIA BARBOSA; CUCIELO, MAIRA SMANIOTTO; SILVEIRA, HENRIQUE SPAULONCI; DOS SANTOS, LUCILENE DELAZARI; PIRES DE CAMPOS ZUCCARI, DEBORA APARECIDA; FERREIRA SEIVA, FABIO RODRIGUES; REITER, RUSSEL J.; DE ALMEIDA CHUFFA, LUIZ GUSTAVO. The proteomic landscape of ovarian cancer cells in response to melatonin. Life Sciences, v. 294, p. 14-pg., 2022-04-01. (19/00906-6)

DE ALMEIDA CHUFFA, LUIZ GUSTAVO; FREIRE, PAULA PACCIELLI; SOUZA, JEFERSON DOS SANTOS; DE MELLO, MARIANA COSTA; NETO, MARIO DE OLIVEIRA; CARVALHO, ROBSON FRANCISCO. ging whole blood transcriptome reveals candidate genes for SARS-CoV-2-related vascular and immune alteration. JOURNAL OF MOLECULAR MEDICINE-JMM, v. 100, n. 2, NOV 2021. (19/00906-6)

CUCIELO, MAIRA SMANIOTTO; CESARIO, ROBERTA CARVALHO; SILVEIRA, HENRIQUE SPAULONCI; GAIOTTE, LETICIA BARBOSA; ALCANTARA DOS SANTOS, SERGIO ALEXANDRE; DE CAMPOS ZUCCARI, DEBORA APARECIDA PIRES; FERREIRA SEIVA, FABIO RODRIGUES; REITER, RUSSEL J.; CHUFFA, LUIZ GUSTAVO DE ALMEIDA. Melatonin Reverses the Warburg-Type Metabolism and Reduces Mitochondrial Membrane Potential of Ovarian Cancer Cells Independent of MT1 Receptor Activation. Molecules, v. 27, n. 14, p. 15-pg., 2022-07-01. (19/00906-6, 18/15797-5)