RESUMO
Zerumbone (ZER) is a phytochemical isolated from plants of the Zingiberaceae family. Numerous studies have demonstrated its diverse pharmacological properties, particularly its potent antitumorigenic activity. This study aimed to assess the antiproliferative effects of ZER on HT-29 cells cultivated in both two-dimensional (2D) monolayer and three-dimensional (3D) spheroid culture systems. The evaluation of growth (size), cell death, and cell cycle arrest in 3D spheroid HT-29 cells was correlated with mRNA expression data. Treatment of 2D cells revealed that ZER exhibited cytotoxicity at concentrations above 30 µM, and an IC50 of 83.54 µM (24-h post-ZER treatment) effectively suppressed cell migration. In the 3D model, ZER induced an increase in spheroid volume over a 72-h period attributed to disaggregation and reconfiguration of characteristic zones. Analysis of cell death demonstrated a significant rise in apoptotic cells after 24 h of ZER treatment, along with cell cycle arrest in the G1 phase. Furthermore, ZER treatment resulted in alterations in mRNA expression, affecting key signaling pathways involved in cell death (BCL2 and BBC3), endoplasmic reticulum stress (ERN1), DNA damage (GADD45A), cell cycle regulation (CDKN1A, NFKB1, MYC, and TP53), and autophagy (BECN1 and SQSTM1). These findings suggested that ZER holds promise as a potential candidate for the development of novel anticancer agents that can modulate crucial cell signaling pathways. Additionally, the use of the 3D culture system proved to be a valuable tool in our investigation.
Assuntos
Antineoplásicos , Sesquiterpenos , Humanos , Células HT29 , Apoptose , Antineoplásicos/farmacologia , Sesquiterpenos/farmacologia , Sesquiterpenos/uso terapêutico , Linhagem Celular Tumoral , RNA MensageiroRESUMO
Curcumin is an antiproliferative phytochemical extracted from Curcuma longa L and which has been studied in preclinical drug screening using cell monolayers and animal models. However, several limitations of these culture systems may be overcome by performing screening with three-dimensional (3-D) cell culture. The aim of this study was to investigate the effects of curcumin on cytotoxicity and genotoxicity as well as spheroid growth using cervical adenocarcinoma HeLa cell spheroids by performing RT-PCR mRNA expression of genes involved in cell death (CASP3, CASP8, CASP9, PARP1, BBC3, BIRC5, BCL2, TNF), autophagy (BECN1, SQSTM1), cell cycle regulation (TP53, C-MYC, NF-kB, CDKN1A, m-TOR, TRAF-2), DNA damage repair (H2AFX, GADD45A, GADD45G), oxidative stress (GPX1), reticulum stress (EIF2AK3, ERN1), and invasion (MMP1, MMP9) was investigated. Curcumin was cytotoxic in a concentration-dependent manner. Curcumin-treated spheroids exhibited lower proliferative recovery and cell proliferation attenuation, as observed in the clonogenic assay. Further, no marked genotoxicity was detected. Curcumin-treated spheroids displayed reduced expression of BECN1 (2.9×), CASP9 (2.1×), and PARP1 (2.1×) mRNA. PARP1 inhibition suggested disruption of essential pathways of proliferation maintenance. Downregulated expression of CASP9 mRNA and unchanged expression of CASP3/8 mRNA suggested caspase-independent cell death, whereas downregulated expression of BECN1 mRNA indicated autophagic disruption. Therefore, curcumin exhibits the potential for drug development with antiproliferative activity to be considered for use in cancers.
Assuntos
Curcumina , Animais , Humanos , Curcumina/farmacologia , Caspase 3 , Células HeLa , Caspases , Proliferação de CélulasRESUMO
Zerumbone (ZER) is a phytochemical with antioxidant and antiproliferative properties. This study evaluated the cytoxicity of ZER combined with chemotherapeutic agents and the expression of mRNA genes related to cell cycle, cell death, xenobiotic metabolism, DNA damage, and endoplasmic reticulum (ER) stress in HepG2/C3A cells. ZER was cytotoxic (IC50, 44.31 µM). ZER-induced apoptosis was related to BBC3 and ERN1 upregulation (ER stress), and its antiproliferative effects were attributable to MYC, IGF1, and NF-kB mRNA inhibition. ZER-induced G2/M arrest and DNA damage was associated with mRNA expression of cell cycle (CDKN1A) and DNA damage (GADD45A) genes. Increased CYP1A2 and CYP2C19 mRNA expression suggested ZER metabolization, and reduced CYP1A1 and CYP2D6 expression indicated a longer time of action of ZER in the cell, enhancing its pharmacological effect. ZER downregulated TP53, PARP1, BIRC5 (apoptosis), and MAP1LC3A (autophagy). In apoptosis assay, the data of the association treatments with ZER suggested antagonism. In cytotoxicity assay, the data of the association treatments with ZER suggested synergism action to cisplatin and antagonism action to doxorubicin and 5-fluorouracil. Thus, ZER has potential for application in chemotherapy as it modulates mRNA targets; however, it may not have the desired efficiency when combined with other chemotherapeutic agents.
Assuntos
Antineoplásicos , Sesquiterpenos , Citocromo P-450 CYP1A2 , Citocromo P-450 CYP2C19 , Cisplatino/farmacologia , Antioxidantes/farmacologia , NF-kappa B , Citocromo P-450 CYP2D6/farmacologia , Citocromo P-450 CYP1A1 , Xenobióticos/farmacologia , Sesquiterpenos/farmacologia , Apoptose , Dano ao DNA , Antineoplásicos/farmacologia , Compostos Fitoquímicos/farmacologia , RNA Mensageiro , Doxorrubicina/farmacologia , Fluoruracila/farmacologia , Linhagem Celular TumoralRESUMO
1α, 25, dihydroxyvitamin D3 (1,25D), the active form of vitamin D3, has antitumor properties in several cancer cell lines in vitro. Salinomycin (Sal) has anticancer activity against cancer cell lines. This study aims to examine the cytotoxic and antiproliferative effect of Sal associated with 1,25D on MCF-7 breast carcinoma cell line cultured in monolayer (2D) and three-dimensional models (mammospheres). We also aim to evaluate the molecular mechanism of Sal and 1,25D-mediated effects. We report that Sal and 1,25D act synergistically in MCF-7 mammospheres and monolayer causing G1 cell cycle arrest, reduction of mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) overproduction with a long-lasting cytotoxic response represented by clonogenic and mammosphere assay. We observed the induction of cell death by apoptosis with upregulation in mRNA levels of apoptosis-related genes (CASP7, CASP9, and BBC3). Extensive cytoplasmic vacuolization, a morphological characteristic found in paraptosis, was also seen and could be triggered by endoplasmic reticulum stress (ER) as we found transcriptional upregulation of genes related to ER stress (ATF6, GADD153, GADD45G, EIF2AK3, and HSPA5). Overall, Sal and 1,25D act synergistically, inhibiting cell proliferation by activating simultaneously multiple death pathways and may be a novel and promising luminal A breast cancer therapy strategy.
Assuntos
Antineoplásicos , Estresse do Retículo Endoplasmático , Antineoplásicos/farmacologia , Apoptose , Técnicas de Cultura de Células em Três Dimensões , Linhagem Celular Tumoral , Colecalciferol/farmacologia , Humanos , Células MCF-7 , PiranosRESUMO
Fluopsin C is an antibiotic compound derived from secondary metabolism of different microorganisms, which possesses antitumor, antibacterial, and antifungal activity. Related to fluopsin C antiproliferative activity, the aim of this study was to examine the following parameters: cytotoxicity, genotoxicity, cell cycle arrest, cell death induction (apoptosis), mitochondrial membrane potential (MMP), colony formation, and mRNA expression of genes involved in adaptive stress responses and cellular death utilizing a monolayer. In addition, a three-dimensional cell culture was used to evaluate the effects on growth of tumor spheroids. Fluopsin C was cytotoxic (1) producing cell division arrest in the G1 phase, (2) elevating expression of mRNA of the CDKN1A gene and (3) decrease in expression of mRNA H2AFX gene. Further, fluopsin C enhanced DNA damage as evidenced by increased expression of mRNA of GADD45A and GPX1 genes, indicating that reactive oxygen species (ROS) may be involved in the observed genotoxic response. Reticulum stress was also detected as noted from activation of the ribonuclease inositol-requiring protein 1 (IRE1) pathway, since a rise in mRNA expression of the ERN1 and TRAF2 genes was observed. During the cell death process, an increase in mRNA expression of the BBC3 gene was noted, indicating participation of this antibiotic in oncotic (ischemic) cell death. Data thus demonstrated for the first time that fluopsin C interferes with the volume of tumor spheroids, in order to attenuate their growth. Our findings show that fluopsin C modulates essential molecular processes in response to stress and cell death.
Assuntos
Apoptose , Dano ao DNA , Antibacterianos/farmacologia , Morte Celular , Humanos , Hidroxilaminas , Células MCF-7 , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND/OBJECTIVE: There is evidence that metabolic profile changes after Roux-Y gastric bypass (RYGB), especially due to modifications in the gastrointestinal tract. In addition, previous studies have suggested that probiotics can modify the microbiome and produce metabolites important for metabolic health maintenance. In this sense, the aim of this study was to verify the influence of probiotic supplementation on the plasma metabolite profile after RYGB. METHODS: This was a randomized, double-blind, placebo-controlled clinical trial conducted with 31 patients subjected to RYGB surgery, randomized in probiotic group that was supplemented with a probiotic supplement (FloraVantage®) for 3 months after surgery or a placebo group. Plasma metabonomics was performed using nuclear magnetic resonance (NMR) at the preoperative period (T0) and at 45-50 days (T1) and 90-95 days (T2) during the postoperative period/intervention. RESULTS: Reductions in trimethylamine-N-oxide (TMAO) and alanine were observed in both groups, however this reduction was greater in the probiotic group (TMAO 13.82%, p = 0.01 and alanine 14.03%, p = 0.03) at T2. Additionally, ß-hydroxybutyrate (BHB) levels increased 10.77% in the probiotic group (p = 0.03) compared to the placebo group at T2. CONCLUSION: Supplementation with Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 was able to associate with significant differences in relevant plasma metabolites associated with improved metabolic health.
Assuntos
Derivação Gástrica , Probióticos , Humanos , Ácido 3-Hidroxibutírico , Estudos Prospectivos , Glicemia/metabolismo , Probióticos/uso terapêutico , Suplementos Nutricionais , Método Duplo-Cego , Alanina , ÓxidosRESUMO
Preclinical studies have shown that diosgenin, a steroidal sapogenin, is a promising phytochemical for treating different pathological conditions, such as cancer, diabetes, and cardiovascular diseases. However, the toxicological safety of this molecule for therapeutic use in humans needs to be better understood. Thus, this study aimed to evaluate the mechanisms of action of diosgenin in HepG2/C3A human hepatocellular carcinoma cells. Cytotoxicity, genotoxicity, alterations in the cell cycle, and cell death (apoptosis) were investigated and associated with the gene expression profile of pathways involved in these processes. The effects of diosgenin on the growth of spheroids were also tested. Diosgenin induced a dose-dependent reduction in cell viability and cell cycle arrest in S and G2/M phases and apoptosis in response to DNA damage. Apoptosis was associated with an increase in the expression of BBC3, a participant in the intrinsic apoptosis pathway. Diosgenin also promoted an increase in volume and greater cellular breakdown in spheroids. These results allowed a better understanding of the toxicity of diosgenin in human cells and contributed to the development of treatments based on this phytochemical.
Assuntos
Carcinoma Hepatocelular , Diosgenina , Neoplasias Hepáticas , Apoptose , Proteínas Reguladoras de Apoptose , Carcinoma Hepatocelular/genética , Comunicação Celular , Diosgenina/farmacologia , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Proteínas Proto-OncogênicasRESUMO
AIM: The use of probiotics as adjuvants in the treatment of eating disorders, known as psychobiotics, has already been investigated as a means of modulating the microbiota-gut-brain axis. This study aimed to assess the effect of probiotic supplementation on binge eating and food addiction in subjects after Roux-en-Y gastric bypass surgery. METHODS: This is a randomized, double-blind, placebo-controlled trial involving 101 patients who received probiotic (Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07) or placebo supplements for 90 days after bariatric surgery, starting on the seventh postoperative day. They were evaluated preoperatively (T0) and postoperatively at 90 days (T1) and 1 year (T2) after surgery. The Yale Food Addiction Scale (YFAS) and Binge Eating Scale (BES) were applied to assess food addiction and binge eating, respectively. RESULTS: Before surgery, one-third of the patients presented with a food addiction and binge eating diagnosis. The number of symptoms of YFAS and the BES score decreased significantly in both groups at T1 compared to T0. However, a significant effect of treatment with probiotics was observed 1 year after surgery (T2). Both the number of symptoms of food addiction and the binge eating score were lower in the probiotic group than in the placebo group (p=0.037 and p=0.030, respectively). CONCLUSION: The use of probiotic supplementation for 90 days in the immediate postoperative period may decrease food addiction symptoms and binge eating score up to 1 year after surgery compared to controls.
Assuntos
Transtorno da Compulsão Alimentar , Bulimia , Dependência de Alimentos , Derivação Gástrica , Probióticos , Transtorno da Compulsão Alimentar/prevenção & controle , Suplementos Nutricionais , Método Duplo-Cego , Dependência de Alimentos/diagnóstico , Humanos , Probióticos/uso terapêuticoRESUMO
BACKGROUND: The use of probiotics as adjuvants in the treatment of eating disorders, known as psychobiotics, has already been investigated as a means of modulating the microbiota-gut-brain axis. AIM: This study aimed to assess the effect of probiotic supplementation on binge eating and food addiction in subjects after Roux-en-Y gastric bypass surgery. METHODS: This is a randomized, double-blind, placebo-controlled trial involving 101 patients who received probiotic (Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07) or placebo supplements for 90 days after bariatric surgery, starting on the seventh postoperative day. They were evaluated preoperatively (T0) and postoperatively at 90 days (T1) and 1 year (T2) after surgery. The Yale Food Addiction Scale (YFAS) and Binge Eating Scale (BES) were applied to assess food addiction and binge eating, respectively. RESULTS: Before surgery, one-third of the patients presented with a food addiction and binge eating diagnosis. The number of symptoms of YFAS and the BES score decreased significantly in both groups at T1 compared to T0. However, a significant effect of treatment with probiotics was observed 1 year after surgery (T2). Both the number of symptoms of food addiction and the binge eating score were lower in the probiotic group than in the placebo group (p=0.037 and p=0.030, respectively). CONCLUSION: The use of probiotic supplementation for 90 days in the immediate postoperative period may decrease food addiction symptoms and binge eating score up to 1 year after surgery compared to controls.
RESUMO - RACIONAL: O uso de probióticos como coadjuvantes no tratamento de distúrbios alimentares, conhecidos como psicobióticos, já foi investigado na modulação do eixo intestino-microbiota-cérebro. OBJETIVO: Analisar a influência da suplementação com probióticos no vício e compulsão alimentar em indivíduos submetidos à cirurgia de bypass gástrico em Y-de-Roux MÉTODOS: Trata-se de um estudo randomizado, duplo-cego, controlado por placebo, envolvendo 101 pacientes que receberam suplementação de probiótico (Lactobacillus acidophilus NCFM e Bifidobacterium lactis Bi-07) ou placebo, durante 90 dias após a cirurgia bariátrica, com início no sétimo dia de pós-operatório. Os indivíduos foram avaliados no pré-operatório (T0) e no pós-operatório aos 90 dias (T1) e 1 ano (T2) após a cirurgia. A Escala de Dependência Alimentar de Yale (YFAS) e a Escala de Compulsão Alimentar Periódica (ECAP) foram aplicadas para avaliar o vício e compulsão alimentar, respectivamente. RESULTADOS: Antes da cirurgia, um terço dos pacientes apresentou diagnóstico de dependência alimentar e compulsão alimentar. O número de sintomas da YFAS e a pontuação da ECAP diminuiu significativamente em ambos os grupos em T1 em comparação com T0. Entretanto, um ano após a cirurgia (T2), tanto o número de sintomas de vício alimentar como a pontuação de compulsão alimentar foram menores no grupo probiótico do que no grupo placebo (p = 0,037 e p = 0,030, respectivamente). CONCLUSÃO: A utilização de suplemento probiótico durante 90 dias após a cirurgia pode diminuir os sintomas de vício alimentar e a pontuação de compulsão alimentar um ano após a cirurgia em comparação com o grupo controle.
RESUMO
BACKGROUND: Obesity is considered a chronic inflammatory disease and transforming growth factor beta 1 (TGFß1) might exert important roles in disease pathogenesis regulating adipocyte differentiation and immune-inflammatory environment. However, the role of this cytokine as a biomarker in obesity is poorly addressed. Therefore, the present study aimed to evaluate the impact of TGFB1 polymorphisms and TGFß1 plasmatic levels in obesity METHODS AND RESULTS: TGFB1 promoter region polymorphisms (rs1800468, G-800A and rs1800469, C-509 T) were evaluated in 75 obese patients and 45 eutrophic patients through PCR-RFLP and plasmatic TGFß1 was quantified through ELISA from 37 of the obese patients, and correlations with clinical and biochemical parameters were tested. Despite no association was found between TGFB1 polymorphisms and obesity susceptibility, several correlations with clinical data were noted. Among others, AC haplotype negatively correlated with plasmatic TGFß1, while plasmatic TGFß1 negatively correlated with C-reactive protein and positively correlated with liver abnormalities on ultrasound and, specifically, with steatosis presence and degree. Conversely, GT haplotype, which associates with higher TGFß1 production, was also positively correlated with the same parameters of liver abnormalities. Further, plasmatic vitamin D negatively correlated with TGFß1, while positively correlated with AC haplotype. CONCLUSION: Overall, the results indicate that TGFß1 might exert important roles in obesity pathophysiology and correlate with biochemical and clinical parameters both at systemic protein as well as at genetic level. Importantly, the consistent positive correlation at both levels with steatosis might suggest this cytokine as a biomarker for this hepatic abnormality in obese patients.