RESUMEN
BACKGROUND: Tumor heterogeneity is a main contributor of resistance to anti-cancer targeted agents though it has proven difficult to study. Unfortunately, model systems to functionally characterize and mechanistically study dynamic responses to treatment across coexisting subpopulations of cancer cells remain a missing need in oncology. METHODS: Using single cell cloning and expansion techniques, we established monoclonal cell subpopulations (MCPs) from a commercially available epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer cell line. We then used this model sensitivity to the EGFR inhibitor osimertinib across coexisting cell populations within the same tumor. Pathway-centered signaling dynamics associated with response to treatment and morphological characteristics of the MCPs were assessed using Reverse Phase Protein Microarray. Signaling nodes differentially activated in MCPs less sensitive to treatment were then pharmacologically inhibited to identify target signaling proteins putatively implicated in promoting drug resistance. RESULTS: MCPs demonstrated highly heterogeneous sensitivities to osimertinib. Cell viability after treatment increased > 20% compared to the parental line in selected MCPs, whereas viability decreased by 75% in other MCPs. Reduced treatment response was detected in MCPs with higher proliferation rates, EGFR L858R expression, activation of EGFR binding partners and downstream signaling molecules, and expression of epithelial-to-mesenchymal transition markers. Levels of activation of EGFR binding partners and MCPs' proliferation rates were also associated with response to c-MET and IGFR inhibitors. CONCLUSIONS: MCPs represent a suitable model system to characterize heterogeneous biomolecular behaviors in preclinical studies and identify and functionally test biological mechanisms associated with resistance to targeted therapeutics.
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Compuestos de Anilina , Antineoplásicos , Resistencia a Antineoplásicos , Receptores ErbB , Transducción de Señal , Humanos , Resistencia a Antineoplásicos/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Antineoplásicos/farmacología , Receptores ErbB/metabolismo , Receptores ErbB/genética , Línea Celular Tumoral , Compuestos de Anilina/farmacología , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/genética , Acrilamidas/farmacología , Supervivencia Celular/efectos de los fármacos , Indoles , PirimidinasRESUMEN
Background and Aim: This study aimed to evaluate the effects of bleaching agents on the fracture resistance of endodontically treated teeth using different intraorifice barrier (IOB) materials. Materials and Methods: The endodontic treatment was performed for 160 mandibular premolars, and then, the teeth were divided into four groups according to the IOB: Ionoseal, Biodentine, ProRoot MTA, and TheraBase. Then, these teeth were subdivided into four subgroups (n = 10) based on the bleaching agents as distilled water (control), hydrogen peroxide 35% (HP), sodium perborate (SP), and carbamide peroxide 37% (CP). The access cavities were restored with composite resin after applying the bleaching agents for 7 days. The fracture resistance test was performed using a universal testing machine. Data were statistically analyzed, and the significance level was set at 5%. A scanning electron microscope was used to evaluate the effect of bleaching agents on the surfaces of IOBs. Results: The highest fracture resistance values were observed in Biodentine groups with significant differences compared to Ionoseal and ProRoot MTA (P <.05). The distilled water groups showed significantly the highest fracture resistance compared to SP and HP groups (P <.05). There was no significant difference between SP, HP, and CP groups (P >.05). It was demonstrated that the morphological surface of the intact IOBs (control) was different from the surface of IOBs treated with bleaching agents. Conclusion: The intracoronal bleaching procedures affected negatively the fracture resistance of the endodontically treated teeth.
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Blanqueadores , Blanqueamiento de Dientes , Diente no Vital , Humanos , Blanqueamiento de Dientes/métodos , Peróxido de Hidrógeno , Peróxido de Carbamida , Resinas Compuestas , AguaRESUMEN
We evaluated the effects of supplementation of L-alanine and L-glutamine on blood glucose levels and biochemical parameters in alloxan-induced diabetic rat. Forty-nine animals were distributed into seven equal groups. Except for the non-diabetic control, diabetes was induced in all groups by intravenous alloxan injection followed by daily supplementation with amino acids for 14 days. Weight and blood glucose were monitored during supplementation, while biochemical parameters such as liver and renal functions, lipid profile, and antioxidant markers were evaluated post-intervention. A significant increase (p < .05) in weight and decrease in blood glucose were observed in the amino acid(s) treated groups. The supplementation with both amino acids restored important tissue antioxidants, liver and kidney functions and rescued islets cells degeneration. Histopathological examinations of important tissues showed the restoration of alloxan-induced physiopathological changes by the amino acids. Thus, these amino acids might serve as nutraceuticals for the management and treatment of diabetes. PRACTICAL APPLICATIONS: The discovery and production of antidiabetic bioactive compounds are often challenging, and the existing antidiabetic drugs are expensive. Amino acids are key regulators of glucose metabolism, insulin secretion, and insulin sensitivity; thus, they can play a crucial role in alleviating diabetes. Here, we present findings that strongly suggest the potential of pure amino acids (L-alanine and L-glutamine) for the management and treatment of diabetes. We show that these amino acids, when supplemented singly or coadministered can lower blood glucose levels and restore several other biochemical parameters implicated in diabetes. Hence, these cheap amino acids may be consumed as nutraceuticals or food supplements by diabetics for the treatment/management of diabetes. Foods rich in these amino acids may also be consumed as part of the diet of diabetic patients.
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Glucemia , Diabetes Mellitus Experimental , Ratas , Animales , Glucemia/metabolismo , Glutamina/efectos adversos , Ratas Wistar , Aloxano/efectos adversos , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismo , Hipoglucemiantes/farmacología , Antioxidantes/uso terapéutico , Alanina/efectos adversosRESUMEN
Targetable alterations in cancer offer novel opportunities to the drug discovery process. However, pre-clinical testing often requires solubilization of these drugs in cosolvents like dimethyl sulfoxide (DMSO). Using a panel of cell lines commonly used for in vitro drug screening and pre-clinical testing, we explored the DMSO off-target effects on functional signaling networks, drug targets, and downstream substrates. Eight Non-Small Cell Lung Cancer (NSCLC) cell lines were incubated with three concentrations of DMSO (0.0008%, 0.002%, and 0.004% v/v) over time. Expression and activation levels of 187 proteins, of which 137 were kinases and downstream substrates, were captured using the Reverse Phase Protein Array (RPPA). The DMSO effect was heterogeneous across cell lines and varied based on concentration, exposure time, and cell line. Of the 187 proteins measured, all were statistically different in at least one comparison at the highest DMSO concentration, followed by 99.5% and 98.9% at lower concentrations. Only 46% of the proteins were found to be statistically different in more than 5 cell lines, indicating heterogeneous response across models. These cell line specific alterations modulate response to in vitro drug screening. Ultra-low DMSO concentrations have broad and heterogeneous effects on targetable signaling proteins. Off-target effects need to be carefully evaluated in pre-clinical drug screening and testing.