Antiproliferative and proapoptotic effects of DODAC/synthetic phosphoethanolamine on hepatocellular carcinoma cells

Abstract

BackgroundCurrent studies have demonstrated that DODAC/PHO-S (Dioctadecyldimethylammonium Chloride/Synthetic phosphoethanolamine) liposomes induces cytotoxicity in Hepa1c1c7 and B16F10 murine tumor cells, with a higher proportion than PHO-S. Therefore, our aim was to evaluate the potential of DODAC/PHO-S to elucidate the mechanism of cell death whereby the liposomes induces cytotoxicity in hepatocellular carcinoma Hepa1c1c7, compared to the PHO-S alone.MethodsLiposomes (DODAC/PHO-S) were prepared by ultrasonication. The cell cycle phases, protein expression and types of cell’s death on Hepa1c1c7 were analyzed by flow cytometry. The internalisation of liposomes, mitochondrial electrical potential and lysosomal stability were also evaluated by confocal laser scanning microscopy.ResultsAfter treatment with liposomes (DODAC/PHO-S), we observed a significant increase in the population of Hepa1c1c7 cells experiencing cell cycle arrest in the S and G2/M phases, and this treatment was significantly more effective to promote cell death by apoptosis. There also was a decrease in the mitochondrial electrical potential; changes in the lysosomes; nuclear fragmentation and catastrophic changes in Hepa1c1c7 cells. The liposomes additionally promoted increases in the expression of DR4 receptor, caspases 3 and 8, cytochrome c, p53, p21, p27 and Bax. There was also a decrease in the expression of Bcl-2, cyclin D1, CD90 and CD44 proteins.ConclusionThe overall results showed that DODAC/PHO-S liposomes were more effective than PHO-S alone, in promoting cytotoxicity Hepa1c1c7 tumor cells, activating the intrinsic and extrinsic pathways of programmed cell death.

Ethanolamine-phosphate phospho-lyase (ETNPPL) contributes to the diagnosis, prognosis, and therapy of hepatocellular carcinoma

“…For example, in renal cell carcinomas, PEA can inhibit lung metastasis of tumor cells, and down-regulate a series of tumor-related proteins such as cyclin D1, vascular endothelial growth factor receptor-1 (VEGFR1), etc . ( Luna et al, 2018 ). PEA induced G2/M cell cycle arrest, abnormal mitochondrial membrane potential, and caspase-3-dependent apoptosis in melanoma cells ( Ferreira et al, 2012 ).…”

2-Aminoethyl Dihydrogen Phosphate (2-AEH2P) Associated with Cell Metabolism-Modulating Drugs Presents a Synergistic and Pro-Apoptotic Effect in an In Vitro Model of the Ascitic Ehrlich Tumor

“…With this aim, our group has expressed interest in exploring the activity of 2-aminoethyl dihydrogen phosphate (2-AEH2P) as a therapeutic adjuvant through in vitro and in vivo studies involving various types of cancer. Indeed, 2-AEH2P has modulated proliferative and apoptotic effects in human breast cancer cells with estrogen, progesterone, and glucocorticoid receptors (MCF-7) [24], in triple-negative breast cancer cells (MDA-MB-231) [25], murine melanoma (B16-F10) [26], K-562 and K-562 MDR(+) myeloid leukemia [27], and murine hepatoma (Hepa-1c1c7) [28], indicating significant data in these cell lines regarding the reduction of mitochondrial membrane potential and the expression of markers involved in cell death, angiogenesis, and proliferation. In turn, paclitaxel (PTX) is a widely used chemotherapy agent in the treatment of advanced metastatic breast cancer [29].…”

Análise crítica dos efeitos biológicos da fosfoetanolamina sintética no câncer

“…Em 28 de fevereiro de 2008, em conjunto com seu companheiro de pesquisa Salvador Claro Neto, e outros pesquisadores, realizaram o depósito da patente de uma nova rota de síntese da pETN na forma sólida com cálcio, magnésio e zinco e na forma de solução com monoetanolamina, com baixo custo de produção (PI 0800460-9 A2). Chierice contava com o Prof. Dr. Durvanei Augusto Maria, do Instituto Butantan, com quem mais publicou trabalhos sobre a pETN (Ferreira et al, 2011;2012a;2012b;2013a;2013b;2013c;Luna et al, 2016;2018a;2018b). Durvanei constatou que o efeito biológico antitumoral da pETN é dependente da concentração (Ferreira et al, 2012;2012b;2013a).…”

“…Em análises com microscopia eletrônica de varredura, foi possível observar os lipossomas aderidos às células cancerígenas (melanoma) antes de serem internalizados, sugerindo uma afinidade e atração eletromagnética [Figura 11a] (Luna et al, 2016;2018a). O estudo de microscopia confocal a laser com o 2-AEH2F marcado com fluorescente Dil mostrou a internalização do composto para o interior das células de carcinomas hepatocelulares [Figura 11b] (Luna et al, 2018b). A 2-AEH2F também mostrou potencial antitumoral em linhagens celulares de adenocarcinoma de mama (Silva, 2016;Bonfim Neto, 2018), de carcinoma espinocelular de língua humana (de Carvalho, 2017) e de leucemia (Conceição, 2017).…”

“…O biobanco possui amostras de tecidos humanos normais e patológicos, análises a níveis teciduais, celulares, subcelulares, entre outras (Uhlen et al, 2010;Karlsson et al, 2021). O motivo da escolha do tecido hepático foi devido aos dados e evidências prévias da internalização da pETN lipossomal para o interior de hepatocarcinomas produzidos em nosso laboratório (Luna et al, 2018b). O gene escolhido para a análise foi o PCYT2 como palavra-chave e critério de correlação entre os pathways.…”