RESUMEN
Considering the rapidly increasing prevalence of obesity worldwide, the number of weight control drugs is very few. Incretin-based therapies are currently being developed to achieve weight control, and Glucagon-Like Peptide-1 Receptor Agonists (GLP-1RA) are used in incretin-based therapies. This study aimed to investigate the cytotoxicity of exenatide, a GLP-1A, on 3T3-L1 adipocytes and the effect of exenatide on the expression of adipogenesis-related genes, insulin and glucose levels, and apoptosis. Cytotoxic activity of exenatide on 3T3-L1 adipocytes was determined by MTT method. Gene expression levels were determined by qPCR. Apoptosis studies were performed on the Muse Cell Analyzer. C1q/TNF-related protein-3 (CTRP3) expression levels were found to be higher in exenatide treated adipocyte cells than in control cells (p < 0.001). Adipocyte cells treated with exenatide were found to have lower PPAR-γ gene expression levels when compared to control adipocyte cells (p < 0.001). Intracellular insulin (p < 0.001) and glucose levels were higher in 3T3-L1 adipocytes treated with exenatide compared to control adipocyte cells. Total apoptosis increased approximately 1.5 times as a result of exenatide administration. The increase in CTRP3 gene expression, which is thought to be a new biomarker for obesity, and the decrease in PPAR-γ gene expression indicate that exenatide is a promising new pharmacotherapeutic agent in the treatment of obesity by regulating the expression of genes related to adipogenesis and lipogenesis and inducing apoptosis.
Asunto(s)
Adipogénesis , Incretinas , Ratones , Animales , Exenatida/farmacología , Exenatida/genética , Exenatida/uso terapéutico , Incretinas/metabolismo , Incretinas/farmacología , Incretinas/uso terapéutico , Alprostadil/metabolismo , Alprostadil/farmacología , Alprostadil/uso terapéutico , Complemento C1q/genética , Complemento C1q/metabolismo , Complemento C1q/farmacología , Receptor del Péptido 1 Similar al Glucagón/genética , Receptor del Péptido 1 Similar al Glucagón/metabolismo , Receptor del Péptido 1 Similar al Glucagón/uso terapéutico , Adipocitos , Células 3T3-L1 , PPAR gamma/metabolismo , Obesidad/genética , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Insulina/farmacología , Insulina/metabolismo , Glucosa/metabolismo , Apoptosis , Expresión Génica , Diferenciación CelularRESUMEN
The ubiquitin-proteasome pathway is an important protein quality control system involved in intracellular homeostasis. To achieve intracellular homeostasis, proteins that are misfolded as a result of translational errors or genetic mutations must be eliminated by the ubiquitin-proteasome pathway. In our previous publications, we determined that 4T1 breast and B16F10 melanoma cancer cells have differential levels of resistance to proteasome inhibitors. Again, in the previous studies, we reported that 4T1 cell cultures, despite being p53-mutant, underwent apoptosis as a result of bortezomib treatment. The first goal of this study was to verify the resistance levels of parental and resistant PC3 prostate cancer cells to bortezomib using WST-1 test. As a result of treatment with different bortezomib concentrations for 48 h, the IC50 value of the parental cells was determined as 32.8 nM and that of the resistant cells was determined as 346 nM. This result showed that the resistant cells were at least 10.5 times more resistant. In addition, to determine whether the resistance gained was reversible or not, the cells were passaged in a medium without bortezomib for one month. The IC50 value determination by WST-1 test showed that the resistant PC3 cells gained an irreversible bortezomib resistance phenotype. The results of the 3D spheroid experiment showed that the 3D spheroid diameter of resistant cells was significantly higher than that of the parental cells. The studies conducted with Western blot showed that ERK1 MAPK T202 phosphorylation and the conversion of autophagy marker LC3-I to LC3-II were significantly increased in parental cells as compared to the resistant cells. Finally, the results showed that while both maternal embryonic leucine zipper kinase (MELK) inhibitor OTSSP167 and Ca2+ chelator BAPTA-AM (also an inhibitor of the expression of antiapoptotic protein GRP78) are promising agents for cancer cells resistant to the proteasome inhibitors, CDK2 inhibitor CVT-313 was found ineffective in both parental and the resistant cells.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis , Autofagia , Biomarcadores de Tumor/metabolismo , Bortezomib/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Neoplasias de la Próstata/patología , Proliferación Celular , Supervivencia Celular , Humanos , Masculino , Células PC-3 , Fosforilación , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/metabolismoRESUMEN
BACKGROUND: The ubiquitin-proteasome pathway is involved in almost all cellular processes (cell cycle, gene transcription and translation, cell survival and apoptosis, cell metabolism and protein quality control) mainly through the specific degradation of the majority of intracellular proteins (>80%) or partial processing of transcription factors (e.g., NF-κB). A growing amount of evidence now indicates that epigenetic changes are also regulated by the ubiquitin-proteasome pathway. Recent studies indicate that epigenetic regulations are equally crucial for almost all biological processes as well as for pathological conditions such as tumorigenesis, as compared to non-epigenetic control mechanisms (i.e., genetic alterations or classical signal transduction pathways). OBJECTIVE: Here, we reviewed the recent work highlighting the interaction of the ubiquitin-proteasome pathway components (e.g., ubiquitin, E1, E2 and E3 enzymes and 26S proteasome) with epigenetic regulators (histone deacetylases, histone acetyltransferases and DNA methyltransferases). RESULTS: Alterations in the regulation of the ubiquitin-proteasome pathway have been discovered in many pathological conditions. For example, a 2- to 32-fold increase in proteasomal activity and/or subunits has been noted in primary breast cancer cells. Although proteasome inhibitors have been successfully applied in the treatment of hematological malignancies (e.g., multiple myeloma), the clinical efficacy of the proteasomal inhibition is limited in solid cancers. Interestingly, recent studies show that the ubiquitin-proteasome and epigenetic pathways intersect in a number of ways through the regulation of epigenetic marks (i.e., acetylation, methylation and ubiquitylation). CONCLUSION: It is therefore believed that novel treatment strategies involving new generation ubiquitinproteasome pathway inhibitors combined with DNA methyltransferase, histone deacetylase or histone acetyltransferase inhibitors may produce more effective results with fewer adverse effects in cancer treatment as compared to standard chemotherapeutics in hematological as well as solid cancers.
Asunto(s)
Epigénesis Genética/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Complejo de la Endopetidasa Proteasomal/metabolismo , Inhibidores de Proteasoma/química , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Ubiquitina/metabolismo , Acetilación , Compuestos de Boro/farmacología , Bortezomib/química , Bortezomib/farmacología , Metilasas de Modificación del ADN/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Glicina/análogos & derivados , Glicina/farmacología , Histona Acetiltransferasas/antagonistas & inhibidores , Histona Desacetilasas/metabolismo , Humanos , Ácidos Hidroxámicos/farmacología , Metilación , FN-kappa B/metabolismo , Inhibidores de Proteasoma/metabolismo , Inhibidores de Proteasoma/farmacología , Transducción de Señal , Compuestos de Terfenilo/farmacología , Ubiquitinación , Ácido Valproico/farmacologíaRESUMEN
The secretion of interleukin (IL)-1 family cytokines is one of the most potent and earliest pro-inflammatory responses triggered by brucellosis. However, the roles of the most recently discovered IL-1 family members, IL-36, IL-37, and IL-38, in the transition into the chronic form of brucellos is remain largely unknown. Therefore, in this study, the roles of IL-36, IL-37, and IL-38 in brucella infections and their effects on the transition from the acute to chronic form of the disease were investigated. Using peripheral blood samples from 40 patients with acute brucellosis, 40 patients with chronic brucellosis, and 40 healthy control subjects, we analysed the serum concentrations of secreted IL-36, IL-37, and IL-38 using ELISA. The findings were confirmed by using RT-qPCR to analyse the mRNA levels of the genes encoding IL-36, IL-37, and IL-38 in peripheral blood mononuclear cells (PBMCs) from 10 randomly selected patients from each of the three groups. Our results showed that serum IL-37 (p < 0.001) and IL-38 (p < 0.001) concentrations were lower in patients with brucellosis than in the healthy controls. In addition, serum IL-37 and IL-38 concentrations were higher in the chronic patient group than in the acute patient group. The mRNA expression levels of IL-37 and IL1F10, genes that encode IL-38, did not affect serum cytokine secretion levels. This result suggests that the high secretion levels of IL-37 and IL-38 may be related to the progression into the chronic form of brucellosis. Our findings will aid in clarifying the mechanism of the transition of brucellosis from the acute to the chronic form of the disease.
Asunto(s)
Brucelosis/sangre , Interleucina-1/sangre , Interleucinas/sangre , Adulto , Células Cultivadas , Enfermedad Crónica , Femenino , Humanos , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Suero/metabolismoRESUMEN
BACKGROUND: The ubiquitin-proteasome pathway is crucial for all cellular processes and is, therefore, a critical target for the investigation and development of novel strategies for cancer treatment. In addition, approximately 30% of newly synthesized proteins never attain their final conformations due to translational errors or defects in post-translational modifications; therefore, they are also rapidly eliminated by the ubiquitin-proteasome pathway. OBJECTIVE: Here, an effort was made to outline the recent findings deciphering the new molecular mechanisms involved in the regulation of ubiquitin-proteasome pathway as well as the resistance mechanisms developed against proteasome inhibitors in cell culture experiments and in the clinical trials. RESULTS: Since cancer cells have higher proliferation rates and are more prone to translational errors, they require the ubiquitin-proteasome pathway for selective advantage and sustained proliferation. Therefore, drugs targeting the ubiquitin-proteasome pathway are promising agents for the treatment of both hematological and solid cancers. CONCLUSION: A number of proteasome inhibitors are approved and used for the treatment of advanced and relapsed multiple myeloma. Unfortunately, drug resistance mechanisms may develop very fast within days of the start of the proteasome inhibitor-treatment either due to the inherent or acquired resistance mechanisms under selective drug pressure. However, a comprehensive understanding of the mechanisms leading to the proteasome inhibitor-resistance will eventually help the design and development of novel strategies involving new drugs and/or drug combinations for the treatment of a number of cancers.