Implications of phenethyl isothiocyanate bioactive compound on the immuno and chemotherapy combination for colon cancer: an in vitro investigation.

• Grant number: 24/06431-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2024
• End date: December 31, 2025
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Guilherme Ribeiro Romualdo
• Grantee: Agnes Suemy Varicoda
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated research grant: 22/06082-8 - Multimodel Drug Screening Platform (MDSP): preclinical and molecular insights for Colon and Liver Cancer management, AP.GR

Abstract

Colon cancer (CC) is one of the most incident neoplasms worldwide, and current therapies do not yield completely satisfactory results. The consumption of a diet rich in bioactive compounds - including isothiocyanates - may reduce the risk of developing CC. Thus, the proposal aims to evaluate whether the administration of phenethyl isothiocyanate (PEITC), abundant in vegetables/fruits, promotes improvement in the antitumor response of combined chemotherapy and immunotherapy with 5-fluorouracil (5-FU)/Bevacizumab in an in vitro model. To this end, HCT-116/CCD-18Co spheroids will be cultured in low-adherence 96-well plates for 48 h. Briefly, cells will be treated with DMEM supplemented with 5-FU plus Bevacizumab or respective vehicles for 24 and 48 h. Spheroids will also be treated with PEITC for 24 and 48 h. Initially, three-dimensional co-cultures will be treated with five different initial concentrations of each therapy or bioactive compound individually to test their effects on viability (MTT) and cytotoxicity (LDH) in their specific models, to calculate the half maximal effective concentration (EC50) of each one. After the individual calculation of EC50 for therapies and bioactive compounds, three-dimensional co-cultures will be treated with the combination of these substances, and concentrations will be 1/5 of the previously calculated EC50. Viability (MTT) and cytotoxicity (LDH) will be evaluated in the three-dimensional models. The viability and cytotoxicity results, using the EC50, will be used for the evaluation of the Combinatorial Index (CI) according to the method proposed by Chou & Talalay (1984) using the CompuSyn software (ComboSyn Inc, USA). CI values of <0.1 will indicate very strong synergy, between 0.1 to 0.3, strong synergy, from 0.3 to 0.7, synergy, from 0.7 to 0.85, moderate synergy, from 0.85 to 0.9, mild synergy, from 0.9 to 1.10, and additive effect.