Toxicological evaluation of diallyl disulfide associated with the chemotherapeutic agent sorafenib: investigation of cell death and expression of proteins in human liver cells in vitro

Author(s)

Ana Rita Thomazela Machado

• Document type: Doctoral Thesis
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2022-05-06

Examining board members

Lusania Maria Greggi Antunes; Maria Sol Brassesco Annichini; Luciana Azevedo; Danielle Palma de Oliveira

• Advisor: Lusania Maria Greggi Antunes

Abstract

Hepatocellular carcinoma is one of the most common cancers in adults. Sorafenib is the drug of choice, but many patients follow tumor progression during chemotherapy which encourage new treatments. The occurrence or development of the disease can be altered by the synergistic effect of traditional drugs and diet bioactive compounds like diallyl disulfide (DADS), and organosulfur isolated from allium with antitumor effects. Thus, the aim of this study was to evaluate the effects of DADS alone and associated with the chemotherapy SORA in cell viability, apoptosis and autophagy, cell migration and invasion, genotoxicity and expression of genes and proteins related to tumor death signaling in hepatocellular carcinoma tumor cells (HepG2). In the isolated treatment, there was a reduction in cell viability from DADS 50 µM and SORA 4 µM. There was inhibition of cell proliferation, increase in the proportion of apoptotic cells and in the percentage of cells in G1/S followed by a decrease in cell migration and invasion in DADS 200 µM and SORA 8 µM. The ratio of autophagic cells and genotoxic damage increased in all treatments tested. There were alterations in the expression of TNF, MMP2, FOS and CHEK2 genes and in the BAX, BCL-2, CASP3, CASP8, LC3 and NRF2 proteins. The DADS and SORA association demonstrated a synergistic effect through the modulation of genes and proteins correlated with cell cycle arrest (decrease in CHEK2), apoptosis (decrease in FOS and increase in BAX) and autophagy (increase of LC3). Our results demonstrated that this association, in simultaneous treatment, can significantly reduce the survival of HepG2 cells by different molecular mechanisms. Since the combination index of this association between SORA and different concentrations of DADS on cell death, this association may represent a promising and alternative therapeutic option in the development of new clinical protocols for the treatment of patients with advanced stage hepatocellular carcinoma. (AU)

• FAPESP's process: 19/08348-2 - Toxicological evaluation of diallyl disulfide associated with the chemotherapeutic agent sorafenib: investigation of cell death and expression of proteins in human liver cells in vitro
• Grantee: Ana Rita Thomazela Machado
• Support Opportunities: Scholarships in Brazil - Doctorate