Carreadores lipídicos nanoestruturados que co-encapsulam partenolídeo e ciclosporina: análises in vitro e in silico

Abstract

Triple-negative breast cancer (TNBC) is considered the most aggressive molecular subtype of breast cancer, associated with the worst prognosis and a high mortality rate. It does not express hormonal receptors or human epidermal growth factor receptor 2, rendering it heterogeneous and primarily treated with chemotherapy. However, the absence of a complete pathological response, low cellular bioavailability and absorption, undesirable resistance and toxicity, and the consequent progression of the disease necessitate new approaches. This study aimed to develop a novel nanoformulation containing Parthenolide (PT) and Cyclosporine (CP) incorporated into nanostructured lipid carriers (NLCs) and to evaluate their effects in TNBC cells. To achieve this, the active compounds were encapsulated in inert lipids using the emulsification-ultrasonication method, followed by physicochemical analyses for quality control and stability assessment. Viability assays were performed using the resazurin reagent, and differentially expressed genes were quantified by qPCR. Human TNBC MDA-MB-231 cells were subjected to seven treatments: PT (non-formulated), CP (non-formulated), PTCP (non formulated combination), NLC (empty lipid carrier), NLC/PT (formulation with only PT), NLC/CP (formulation with only CP), and NLC/PT-CP (formulated combination) for 24, 48, 72, and 168 hours, with cytotoxic effects of the active compounds in the carriers observed only after 168 hours. The encapsulated compounds proved to be more cytotoxic than their free counterparts, with NLC/PT-CP being the most cytotoxic and selective formulation compared to the effects observed in MCF-10A cells (non-tumorigenic human mammary cells). Additionally, 4T1 cells (murine TNBC) were also sensitive to NLC/PT-CP. In MDA-MB-231 cells, NLC/PT CP significantly increased the transcripts of ANXA1, IKBKG, TP53, IL-6, and STAT3. At the protein level, there was a significant increase in the expression of Annexin A1(ANXA1) and p53 in the cytoplasm and a downregulation in IKBKG. Interestingly, the formulation reduced ANXA1 secretion to the extracellular medium. Therefore, we suggest that the mechanism of action of NLC/PT-CP in TNBC cells involves the modulation of ANXA1 endocrine signaling, being promising for the treatment of these tumors.