Melatonin and MT1 and MT2 receptors: effects on apoptosis, proliferation, migration potential of Ovarian Carcinoma cells (SKOV-3)

• Grant number: 18/15797-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2019
• End date: April 30, 2022
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Luiz Gustavo de Almeida Chuffa
• Grantee: Maira Smaniotto Cucielo
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

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, cyt C, and cleaved caspase-3. In parallel, mitochondria will be isolated from SKOV-3 cells to effectively access the membrane potential, energy metabolism, and expression of proteins involved with endogenous melatonin synthesis (AA-NAT and ASMT). These findings may contribute to the current discussion on the role of melatonin in the altered metabolism of neoplastic OC cells. (AU)

Scientific publications

(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)

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)

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)