Abstract:Hepatocellular carcinoma (HCC) is the third most common cause of death related to cancer diseases worldwide. The current treatment options have many limitations and reduced success rates. In this regard, advances in gene therapy have shown promising results in novel therapeutic strategies. However, the success of gene therapy depends on the efficient and specific delivery of genetic material into target cells. In this regard, the main goal of this work was to develop a new lipid-based nanosystem formulation co… Show more
“…For this purpose, the HNP, loaded with the fluorescent dyes coumarin-6 and nile red, and prepared with or without the GalNAc cluster, were incubated with two HCC cell lines (HepG2 and Hep3B) and one non-liver cancer cell line (DBTRG-05MG glioblastoma cell line) at 4°C (for cell binding) or at 37°C (for cell uptake). 31 , 50 After incubation, cells were analysed by flow cytometry ( Figure 5A , B and E ) and confocal microscopy ( Figure 5C and D ). Figure 5 Evaluation of cell binding, uptake and specificity of hybrid nanosystems.…”
Section: Resultsmentioning
confidence: 99%
“…27 Several studies have shown that the use of specific ligands for this receptor allows the efficient and specific targeting of nanosystems to HCC models. 28,30,31 In this regard, Ionis Pharmaceuticals Inc. developed a triantennary N-acetylgalactosamine (GalNAc) cluster that has a high affinity for the asialoglycoprotein receptor. 32 In this study, we designed a novel delivery nanosystem, composed of PLGA nanoparticles coated with a lipid bilayer containing DSPE-PEG (2000)-GalNAc, to mediate a targeted therapeutic strategy to HCC involving the combination of selumetinib and sorafenib.…”
Background: Hepatocellular carcinoma (HCC) is one of the main causes of cancer-related death. Sorafenib, which is the first-line therapy for this disease, is associated with reduced therapeutic efficacy that could potentially be overcome by combination with selumetinib. In this context, the main goal of this work was to develop a new nanosystem, composed of a polymeric core coated by a lipid bilayer containing the targeting ligand GalNAc, to specifically and efficiently co-deliver both drugs into HCC cells, in order to significantly increase their therapeutic efficacy. Methods: The physicochemical characterization of hybrid nanosystems (HNP) and their components was performed by dynamic light scattering, zeta potential, matrix-assisted laser desorption ionization -time of flight mass spectroscopy, and transmission electron microscopy. Cellular binding, uptake and specificity of HNP were evaluated through flow cytometry and confocal microscopy. The therapeutic activity was evaluated namely through: cell viability by the Alamar Blue assay; cell death by flow cytometry using FITC-Annexin V; caspases activity by luminescence; mitochondrial membrane potential by flow cytometry; and molecular target levels by Western blot. Results: The obtained data show that these hybrid nanosystems present high stability and loading capacity of both drugs, and suitable physicochemical properties, namely in terms of size and surface charge. Moreover, the generated formulation allows to circumvent drug resistance and presents high specificity, promoting great cell death levels in HCC cells, but not in non-tumor cells. This potentiation of the antitumor effect of co-loaded drugs was carried out by an increased programmed cell death, being associated with a strong reduction in the mitochondrial membrane potential, a significant increase in the activity of caspases 3/7 and caspase 9, and much greater number of annexin V-positive cells.
Conclusion:The developed formulation resulted in a high and synergistic antitumor effect, revealing a translational potential to improve therapeutic approaches against HCC.
“…For this purpose, the HNP, loaded with the fluorescent dyes coumarin-6 and nile red, and prepared with or without the GalNAc cluster, were incubated with two HCC cell lines (HepG2 and Hep3B) and one non-liver cancer cell line (DBTRG-05MG glioblastoma cell line) at 4°C (for cell binding) or at 37°C (for cell uptake). 31 , 50 After incubation, cells were analysed by flow cytometry ( Figure 5A , B and E ) and confocal microscopy ( Figure 5C and D ). Figure 5 Evaluation of cell binding, uptake and specificity of hybrid nanosystems.…”
Section: Resultsmentioning
confidence: 99%
“…27 Several studies have shown that the use of specific ligands for this receptor allows the efficient and specific targeting of nanosystems to HCC models. 28,30,31 In this regard, Ionis Pharmaceuticals Inc. developed a triantennary N-acetylgalactosamine (GalNAc) cluster that has a high affinity for the asialoglycoprotein receptor. 32 In this study, we designed a novel delivery nanosystem, composed of PLGA nanoparticles coated with a lipid bilayer containing DSPE-PEG (2000)-GalNAc, to mediate a targeted therapeutic strategy to HCC involving the combination of selumetinib and sorafenib.…”
Background: Hepatocellular carcinoma (HCC) is one of the main causes of cancer-related death. Sorafenib, which is the first-line therapy for this disease, is associated with reduced therapeutic efficacy that could potentially be overcome by combination with selumetinib. In this context, the main goal of this work was to develop a new nanosystem, composed of a polymeric core coated by a lipid bilayer containing the targeting ligand GalNAc, to specifically and efficiently co-deliver both drugs into HCC cells, in order to significantly increase their therapeutic efficacy. Methods: The physicochemical characterization of hybrid nanosystems (HNP) and their components was performed by dynamic light scattering, zeta potential, matrix-assisted laser desorption ionization -time of flight mass spectroscopy, and transmission electron microscopy. Cellular binding, uptake and specificity of HNP were evaluated through flow cytometry and confocal microscopy. The therapeutic activity was evaluated namely through: cell viability by the Alamar Blue assay; cell death by flow cytometry using FITC-Annexin V; caspases activity by luminescence; mitochondrial membrane potential by flow cytometry; and molecular target levels by Western blot. Results: The obtained data show that these hybrid nanosystems present high stability and loading capacity of both drugs, and suitable physicochemical properties, namely in terms of size and surface charge. Moreover, the generated formulation allows to circumvent drug resistance and presents high specificity, promoting great cell death levels in HCC cells, but not in non-tumor cells. This potentiation of the antitumor effect of co-loaded drugs was carried out by an increased programmed cell death, being associated with a strong reduction in the mitochondrial membrane potential, a significant increase in the activity of caspases 3/7 and caspase 9, and much greater number of annexin V-positive cells.
Conclusion:The developed formulation resulted in a high and synergistic antitumor effect, revealing a translational potential to improve therapeutic approaches against HCC.
“…U87, A172, H4, U373, U118 cells (1×10 4 cells/well), and HMC3 cells (2×10 4 cells/well) were seeded in 96-well plates, 24 h before treatment. Cells were treated with a range of Roy concentrations (1 to 64 μM), with the vehicle control (DMSO), or with positive control (TMZ -1 to 100 μM), and further incubated for 48 h. Metabolic activity was assessed through Alamar blue ® assay [21]. Briefly, a solution of DMEM-HG medium with 10% (v/v) of a resazurin salt dye stock solution (Sigma.…”
Glioblastoma (GB) is the most aggressive and prevalent glioma within the Central Nervous System. GB exhibit a dismal 5-year survival rate (~6%) due to unfavorable prognosis and lack of viable treatment options. Therefore, novel therapies with plant-derived compounds emerge as very promising. This study aims to investigate the antitumor activity of 7α-acetoxy-6β-hydroxyroyleanone (Roy) in GB cell lines. Roy was isolated from Plectranthus hadiensis Schweinf. and its antitumor mechanism was assessed in a panel of five GB cell lines to mimic tumor heterogeneity. Metabolic activity was evaluated by Alamar Blue® assay, while cell death, cell cycle regulation, mitochondrial membrane potential, and activated caspase-3 were evaluated by flow cytometry. mRNA levels of target genes were measured by qPCR and protein expression was assessed by Western blotting. Cell uptake and impact on mitochondrial morphology were evaluated by confocal microscopy. Roy induced G2/M cell cycle arrest, mitochondrial fragmentation, and apoptosis by inhibiting the expression of anti-apoptotic proteins and increasing activated caspase-3 levels with concentration 6-9 fold lower than those of temozolomide, the first-line treatment. Roy demonstrated a robust antitumor activity against GB cells, without affecting non tumoral cells (microglia and astrocytes), being a promising natural lead compound for the development of novel chemotherapeutic approaches.
“…Human cell lines were seeded in 96-well plates at a density of 5.0 × 10 3 cells/well (HCT116, MCF-7, and A375), and allowed to adhere for 24 h. Cells were treated with serial dilutions of compounds (ranging from 0.1 μmol/L to 50 μmol/L), for an additional 48 h. Effect on cell proliferation was measured by sulforhodamine B (SRB) assay, as described by Raimundo et al [58] and the concentration of compound that induces 50% growth inhibition (GI 50 ) was then determined for each cell line using the GraphPad Prism software version 7.0 (La Jolla, CA, USA).…”
Section: Anticancer Activitymentioning
confidence: 99%
“…Based on the GI 50 values obtained by SRB assay [58], a promising antiproliferative activity against the aforementioned cancer cells has been clearly attested for compound 11. In fact, GI 50 values for derivative 11 range from 3.55 µmol/L to 11.5 µmol/L on the tested cells, significantly lower than those observed for the remaining derivatives (Table 2).…”
Aim: This study discloses the synthesis and the antimicrobial and anticancer activities of four molecules of structural basis saccharin-thiadiazolyl (4), saccharin-pyridyl (6, 8), and tetrazole-thiadiazolyl (11). Methods: Antimicrobial properties of the molecules were evaluated by the well-diffusion method, against Gram-positive bacteria [Staphylococcus aureus American Type Culture Collection (ATCC) 25923, Staphylococcus epidermidis ATCC 12228, Mycobacterium smegmatis ATCC 607], Gram-negative bacteria (Pseudomonas aeruginosa ATCC 27853) and yeast (Saccharomyces cerevisiae ATCC 2601 and Candida albicans ATCC 10231) strains. The anticancer activity of the compounds was assessed through i) proliferation assays for HCT116, MCF-7, and A375 human cell lines [cells were treated with serial dilutions of compounds and the effect on cell propagation was evaluated by sulforhodamine B (SRB) assay]; ii) antiproliferative and cytotoxic assays for glioma-type cell lines A172 (glioblastoma), U87 (brain-likely glioblastoma), and H4 (neuroglioma; cells were treated with diverse concentrations and the cell viability was assessed using a modified Alamar blue® assay). Results: Compound 11 exhibited significant inhibitory activity against S. aureus and S. epidermidis, with the further molecules demonstrating some inhibitory potential against all the tested Gram-positive, Gram-negative, and yeast strains. Similarly, derivative 11 showed an interesting antiproliferative activity against human colon adenocarcinoma (HCT116), human breast adenocarcinoma (MCF-7), and melanoma (A375) cells, with 50% growth inhibition (GI50) values varying from 3.55 µmol/L to 11.5 µmol/L, in the same order of magnitude of those shown by etoposide. Treatment of brain-like glioblastoma cells (U87) with 11, at the concentration of 100 µg/mL, induced a decrease in cell viability by 50% after 48 h and 72 h. Besides, results attained for A172 cells have shown that compound 11 only induces a significant decrease in cell viability upon treatment at 100 µg/mL for 72 h. A divergent observation was recorded for H4 cells, where the treatment with derivative 11 had promoted a significant decrease in cell viability (< 40–60%), even at concentrations as low as 0.39 µg/mL, after 24 h. Conclusions: This investigation reveals the potential of distinct azole-based conjugates, in particular the tetrazole-thiadiazolyl (11) derivative, as scaffolds worth further investigations, in the frame of antimicrobial and antineoplastic chemotherapy.
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