2-Methoxyestradiol-3,17-O,O-bis-sulfamate (STX140) inhibits proliferation and invasion via senescence pathway induction in human BRAFi-resistant melanoma cells

The endogenous estradiol derivative 2-Methoxyestradiol (2-ME) has shown good and wide anticancer activity but suffers from poor oral bioavailability and extensive metabolic conjugation. However, its sulfamoylated derivative, 2-methoxyestradiol-3,17-O,O-bis-sulfamate (STX140), has superior potential as a therapeutic agent, acts by disrupting microtubule polymerization, leading to cell cycle arrest and apoptosis in cancer cells and possesses much better pharmaceutical properties. This study investigated the antiproliferative and anti-invasive activities of STX140 in both SKMEL-28 naïve melanoma (SKMEL28-P) cells and resistant melanoma cells (SKMEL-28R). STX140 inhibited cell proliferation in the nanomolar range while having a less pronounced effect on human melanocytes. Additionally, STX140 induced cell cycle arrest in the G2/M phase and sub-G1, reduced migration, and clonogenic potential in monolayer models, and inhibited invasion in a 3D human skin model with melanoma cells. Furthermore, STX140 induced senescence features in melanoma and activated the senescence machinery by upregulating the expression of senescence genes and proteins related to senescence signaling. These findings suggest that STX140 may hold potential as a therapeutic agent for melanoma treatment

14-weeks combined exercise epigenetically modulated 118 genes of menopausal women with prediabetes

Background: Pre-diabetes precedes Diabetes Mellitus (DM) disease and is a critical period for hyperglycemia treatment, especially for menopausal women, considering all metabolic alterations due to hormonal changes. Recently, the literature has demonstrated the role of physical exercise in epigenetic reprogramming to modulate the gene expression patterns of metabolic conditions, such as hyperglycemia, and prevent DM development. In the present study, we hypothesized that physical exercise training could modify the epigenetic patterns of women with poor glycemic control. Methods: 48 post-menopause women aged 60.3 ± 4.5 years were divided according to their fasting blood glucose levels into two groups: Prediabetes Group, PG (n=24), and Normal Glucose Group, NGG (n=24). All participants performed 14 weeks of physical exercise three times a week. The Infinium Methylation EPIC BeadChip measured the participants’ Different Methylated Regions (DMRs). Results: Before the intervention, the PG group had 12 DMRs compared to NGG. After the intervention, five DMRs remained different. Interestingly, when comparing the PG group before and after training, 118 DMRs were found. The enrichment analysis revealed that the genes were related to different biological functions such as energy metabolism, cell differentiation, and tumor suppression. Conclusion: Physical exercise is a relevant alternative in treating hyperglycemia and preventing DM in post-menopause women with poor glycemic control

Peroxiredoxin 2 in the modulation of the invasive phenotype in NRASmutated melanoma

O melanoma é o câncer de pele mais letal, com taxa de mortalidade de aproximadamente 20% dentre todos os novos casos diagnosticados anualmente. Os principais fatores de risco incluem a exposição à radiação ultravioleta e a predisposição genética. O acúmulo de mutações em nevos melanocíticos predispõe ao desenvolvimento do tumor, sendo a mutação mais comum em BRAF (B-Raf Proto-Oncogene, Serina/Treonina Quinase), presente em cerca de 50% dos casos, seguida pela mutação em NRAS (Homólogo de oncogene viral RAS de neuroblastoma), que representa cerca de 20% dos casos e é associada a um pior prognóstico devido ao aumento das taxas de invasão e metástases. Infelizmente, ainda não há terapias direcionadas para o melanoma com mutação em NRAS, como já existe para pacientes com a mutação em BRAF, que possuem a terapia alvo dirigida. Em busca de possíveis alvos terapêuticos, há estudos relatando a ação de enzimas antioxidantes, como as peroxirredoxinas (PRDX), alterando a homeostase redox intracelular e a tumorigênese. A isoforma PRDX2 já foi estudada em outros tipos de cânceres, pois é capaz de reduzir peróxidos na presença de NADPH pelo acoplamento ao sistema tiorredoxina/tiorredoxina redutase. Curiosamente, a PRDX2 é pouco expressa em melanoma metastático e possui a capacidade de induzir a estabilização de junções aderentes em melanoma com mutação em BRAF. Desse modo, o objetivo do trabalho foi investigar o papel da PRDX2 em melanoma com mutação em NRAS como possível biomarcador e/ou alvo terapêutico. Foi realizado o silenciamento (shRNA) de PRDX2 e o uso de um mimético farmacológico de PRDX2 (Gliotoxina) para verificar seu potencial na proliferação, migração e invasão celular. Além disso, foram feitas análises in silico em tumores de melanoma primário, comparando a expressão da PRDX2 com marcadores e características clínicas relacionadas à progressão do tumor. Os resultados obtidos demonstraram que PRDX2 está mais expressa em melanoma proliferativo do que em melanoma invasivo, e está correlacionada negativamente com marcadores de transição epitélio-mesênquima (N-caderina, Snail, Smad) e regulado positivamente com marcadores de estabilização celular e proliferação (MITF, β-catenina, Queratina). Em experimentos com a gliotoxina, observou-se a capacidade de reduzir a migração em células de melanoma NRAS mutante. As células de melanoma NRAS mutante com baixa expressão de PRDX2 apresentaram sítios de invasão na pele humana reconstruída, confirmando seu potencial invasivo. Os dados in silico de pacientes com melanoma primário com maior expressão PRDX2, apresentaram menor estadiamento do tumor e aumento de sobrevida. Portanto, a PRDX2 pode ser um possível biomarcador e/ou alvo terapêutico para pacientes com melanoma NRAS mutante invasivo.The melanoma is the deadliest skin cancer, with a mortality rate of approximately 20% among all new cases diagnosed annually. The main risk factors include exposure to ultraviolet radiation and genetic predisposition. The accumulation of mutations in melanocytic nevi predisposes tumor development, with the most common mutation in BRAF (B-Raf Proto-Oncogene, Serine/Threonine Kinase), present in about 50% of cases, followed by mutation in NRAS (NRAS Proto-Oncogene, GTPase), which represents about 20% of cases and is associated with worse prognosis due to increased rates of invasion and metastases. Unfortunately, there are still no targeted therapies for NRAS-mutated melanoma, as there already is for patients with BRAFmutated melanoma. In search of possible therapeutic targets, there are already studies reporting the action of antioxidant enzymes, such as peroxiredoxins (PRDX), altering intracellular redox homeostasis and tumorigenesis. The PRDX2 isoform has already been studied in other types of cancers, as it is capable of reducing peroxides in the presence of NADPH by coupling to the thioredoxin/thioredoxin reductase system. Interestingly, PRDX2 is poorly expressed in metastatic melanoma and has the ability to induce stabilization of adherent junctions in BRAFmutated melanoma. Thus, the aim of this study was to investigate the role of PRDX2 in NRASmutated melanoma as a possible biomarker and/or therapeutic target. Gene editing (shRNA) was performed to silence PRDX2 and the use of a pharmacological mimetic of PRDX2 (Gliotoxin) to verify its potential in cell proliferation, migration and invasion. In addition, in silico analyses were performed on primary melanoma tumors, comparing PRDX2 expression with markers and clinical characteristics related to tumor progression. The results obtained demonstrated that PRDX2 is more expressed in proliferative melanoma than in invasive melanoma, is negatively correlated with markers of epithelial-mesenchymal transition (Ncadherin, Snail, Smad) and positively regulated with markers of cell stabilization and proliferation (MITF, β-catenin, Keratin). In experiments with gliotoxin, the ability to reduce migration in NRAS-mutated melanoma cells was observed. NRAS-mutated melanoma cells with low expression of PRDX2 showed invasion sites in reconstructed human skin, confirming their invasive potential. In silico data from patients with primary melanoma with higher PRDX2 expression showed lower tumor staging and increased survival. Therefore, PRDX2 may be a possible biomarker and/or therapeutic target for patients with invasive NRAS-mutated melanoma

14-Week exercise training modifies the DNA methylation levels at gene sites in non-Alzheimer's disease women aged 50 to 70 years

Exercise training emerges as a key strategy in lifestyle modification, capable of reducing the risk of developing Alzheimer's disease (AD) due to risk factors such as age, family history, genetics and low level of education associated with AD. We aim to analyze the effect of a 14-week combined exercise training (CT) on the methylation of genes associated with AD in non-alzheimer's disease women. CT sessions lasted 60 min, occurring three times a week for 14 weeks. Forty non-Alzheimer's disease women aged 50 to 70 years (60.7 ± 4.1 years) with a mean height of 1.6 ± 0.1 m, mean weight of 73.12 ± 9.0 kg and a mean body mass index of 29.69 ± 3.5 kg/m2, underwent two physical assessments: pre and post the 14 weeks. DNA methylation assays utilized the EPIC Infinium Methylation BeadChip from Illumina. We observed that 14 weeks of CT led to reductions in systolic (p = 0.001) and diastolic (p = 0.017) blood pressure and improved motor skills post-intervention. Among 25 genes linked to AD, CT induced differentially methylated sites in 12 genes, predominantly showing hypomethylated sites (negative β values). Interestingly, despite hypomethylated sites, some genes exhibited hypermethylated sites (positive β values), such as ABCA7, BDNF, and WWOX. A 14-week CT regimen was adequate to induce differential methylation in 12 CE-related genes in healthy older women, alongside improvements in motor skills and blood pressure. In conclusion, this study suggest that combined training can be a strategy to improve physical fitness in older individuals, especially able to induce methylation alterations in genes sites related to development of AD. It is important to highlight that training should act as protective factor in older adults.</p