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Unsaturated fatty acids (UFA) are crucial for T-cell effector functions, as they can affect the growth, differentiation, survival, and function of T cells. Nonetheless, the mechanisms by which UFA affects T-cell behavior are ill-defined. Therefore, we analyzed the processing of oleic acid, a prominent UFA abundantly present in blood, adipocytes, and the fat pads surrounding lymph nodes, in CD4+ T cells. We found that exogenous oleic acid increases proliferation and enhances the calcium flux response upon CD3/CD28 activation. By using a variety of techniques, we found that the incorporation of oleic acid into membrane lipids, rather than regulation of cellular metabolism or TCR expression, is essential for its effects on CD4+ T cells. These results provide novel insights into the mechanism through which exogenous oleic acid enhances CD4+ T-cell function.
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The transcription factors (TFs) MyoCD (myocardin) and Elk-1 (ETS Like-1 protein) competitively bind to SRF (serum response factor) and control myogenic- and mitogenic-related gene expression in smooth muscle, respectively. Their functions are therefore mutually inhibitory, which results in a contractile-versus-proliferative phenotype dichotomy. Airway smooth muscle cell (ASMC) phenotype alterations occur in various inflammatory airway diseases, promoting pathological remodeling and contributing to airflow obstruction. We characterized MyoCD and Elk-1 interactions and their roles in phenotype determination in human ASMCs. MyoCD overexpression in ASMCs increased smooth muscle gene expression, force generation, and partially restored the loss of smooth muscle protein associated with prolonged culturing while inhibiting Elk-1 transcriptional activities and proliferation induced by EGF (epidermal growth factor). However, MyoCD overexpression failed to suppress these responses induced by FBS, as FBS also upregulated SRF expression to a degree that allowed unopposed function of both TFs. Inhibition of the RhoA pathway reversed said SRF changes, allowing inhibition of Elk-1 by MyoCD overexpression and suppressing FBS-mediated contractile protein gene upregulation. Our study confirmed that MyoCD in increased abundance can competitively inhibit Elk-1 function. However, SRF upregulation permits a dual contractile-proliferative ASMC phenotype that is anticipated to exacerbate pathological alterations, whereas therapies targeting SRF may inhibit pathological ASMC proliferation and contractile protein gene expression.
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Proliferação de Células , Contração Muscular , Miócitos de Músculo Liso , Proteínas Nucleares , Fenótipo , Fator de Resposta Sérica , Transativadores , Proteínas Elk-1 do Domínio ets , Proteína rhoA de Ligação ao GTP , Humanos , Fator de Resposta Sérica/metabolismo , Fator de Resposta Sérica/genética , Proteínas Elk-1 do Domínio ets/metabolismo , Proteínas Elk-1 do Domínio ets/genética , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Proteína rhoA de Ligação ao GTP/metabolismo , Transativadores/metabolismo , Transativadores/genética , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Células Cultivadas , Regulação da Expressão Gênica , Transdução de Sinais , Fator de Crescimento Epidérmico/metabolismoRESUMO
PURPOSE: To establish reference ranges (RRs) for stimulation index of T cell proliferation triggered by phytohemagglutinin (PHA-SI) and Bacillus Calmette-Guérin (BCG-SI). METHODS: This study investigated data from 359 healthy children and 35 patients with cellular immunodeficiency as positive controls (2010-2021). We applied a colorimetric-based method (BrdU) to measure proliferation and determine the RRs at the 2.5th and 97.5th percentiles (95% confidence intervals). A cross-validation approach was performed. RESULTS: In healthy controls, the RRs for PHA-SI and BCG-SI ranged between 3 and 5.2 and 2.52 to 5.2, respectively. PHA-SI and BCG-SI were in Severe Combined Immunodeficiency (SCID) patients from 1.2 to 2.5 and 0 to 2, while in Mendelian susceptibility to mycobacterial diseases (MSMD) patients, 2.53 to 4.5 and 0.74 to 2.2, respectively. The thresholds' accuracy was checked for testing reference intervals with diagnostic effects. CONCLUSION: This study establishes PHA-SI and BCG-SI reference ranges to aid in diagnosing and treating congenital immunodeficiency diseases.
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Vacina BCG , Mycobacterium bovis , Criança , Humanos , Irã (Geográfico) , Fito-Hemaglutininas/farmacologia , Valores de Referência , LinfócitosRESUMO
We studied the effect of human lactoferrin on cells of the hippocampal dentate gyrus of 2-2.5-month-old male C57BL/6 mice after acute gamma irradiation of the head in a dose of 8 Gy from a 60Co source. Immediately after irradiation some animals received an intraperitoneal injection of human lactoferrin (4 mg/mouse). The appearance of TUNEL+ cells in the subgranular zone 6 h after irradiation was accompanied by a corresponding decrease in the number of Ki-67- and DCX-immunoreactive cells. Administration of lactoferrin had a protective effect on mouse brain cells, which manifested in a decrease in the number of TUNEL+ cells (by 77% relative to the irradiation alone) and an increase in the number of proliferating cells (from 16 to 61% relative to control animals) and immature neurons (from 14 to 22% relative to control animals) in the dentate gyrus of the hippocampus.
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Giro Denteado , Lactoferrina , Humanos , Camundongos , Masculino , Animais , Lactente , Lactoferrina/farmacologia , Proteína Duplacortina , Camundongos Endogâmicos C57BL , Hipocampo , Encéfalo , Neurogênese/fisiologia , Proliferação de CélulasRESUMO
BACKGROUND: Neuroblastoma is a common pediatric malignancy with poor survival for high-risk disease. Mesenchymal stromal cells (MSCs) have innate tumor-homing properties, enabling them to serve as a cellular delivery vehicle, but MSCs have demonstrated variable effects on tumor growth. We compared how placental MSCs (PMSCs) and bone marrow-derived MSCs (BM-MSCs) affect proliferation of neuroblastoma (NB) cells in vitro. METHODS: Indirect co-culture assessed proliferative effects of 18 MSCs (early-gestation PMSCs (n = 9), term PMSCs (n = 5), BM-MSCs (n = 4) on three high-risk NB cell lines (NB1643, SH-SY5Y, and CHLA90). Controls were NB cells cultured in media alone. Proliferation was assessed using MTS assay and measured by fold change (fc) over controls. PMSCs were sub-grouped by neuroprotective effect: strong (n = 7), intermediate (n = 3), and weak (n = 4). The relationship between MSC type, PMSC neuroprotection, and PMSC gestational age on NB cell proliferation was assessed. RESULTS: NB cell proliferation varied between MSC groups. BM-MSCs demonstrated lower proliferative effects than PMSCs (fc 1.18 vs 1.44, p < 0.001). Neither gestational age nor neuroprotection significantly predicted degree of proliferation. Proliferative effects of MSCs varied among NB cell lines. BM-MSCs had less effect on CHLA90 (fc 1.01) compared to NB1643 (fc 1.33) and SH-SY5Y (fc 1.20). Only NB1643 showed a difference between early and term PMSCs (p = 0.04). CONCLUSION: Effects of MSCs on NB cell proliferation vary by MSC source and NB cell line. BM-MSCs demonstrated lower proliferative effects than most PMSCs. MSC neuroprotection was not correlated with proliferation. Improved understanding of MSC proliferation-promoting mechanisms may provide valuable insight into selection of cells best suited as drug delivery vehicles. LEVEL OF EVIDENCE: N/A. TYPE OF STUDY: Original Research.
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Proliferação de Células , Técnicas de Cocultura , Células-Tronco Mesenquimais , Neuroblastoma , Placenta , Humanos , Neuroblastoma/patologia , Linhagem Celular Tumoral , Feminino , Placenta/citologia , Gravidez , Células da Medula Óssea , Idade GestacionalRESUMO
INTRODUCTION: Statins have demonstrated chondroprotective effects by reducing inflammation and mitigating extracellular matrix degradation. However, statins are also reported to be cytotoxic to several types of cells. Early-onset osteoarthritis (OA) is characterized by synovial inflammation, which adversely affects hyaluronan (HA) production in fibroblast-like synoviocytes (FLSs). Nevertheless, the precise effects of statins on the synovium remain unclear. METHODS: This study investigated the impact of lovastatin on human FLSs, and HA secretion-related genes, signaling pathways, and production were evaluated. RESULTS: The findings revealed that high doses of lovastatin (20 or 40 µM) decreased FLS viability and increased cell death. FLS proliferation ceased when cultured in a medium containing 5 or 10 µM lovastatin. mRNA expression analysis demonstrated that lovastatin (5 and 10 µM) upregulated the gene level of hyaluronan synthase 1 (HAS1), HAS2, and proteoglycan 4 (PRG4), but not HAS3. While the expression of multidrug resistance-associated protein 5 transporter gene remained unaffected, both inward-rectifying potassium channel and acid-sensing ion channel 3 were upregulated. Western blot further confirmed that lovastatin increased the production of HAS1 and PRG4, and activated the PKC-α, ERK1/2, and p38-MAPK signaling pathways. Additionally, lovastatin elevated intracellular cAMP levels and HA production in FLSs. CONCLUSION: Lovastatin impairs cellular proliferation but enhances HA production in human FLSs.
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Inibidores de Hidroximetilglutaril-CoA Redutases , Sinoviócitos , Humanos , Sinoviócitos/metabolismo , Ácido Hialurônico/metabolismo , Lovastatina/farmacologia , Lovastatina/metabolismo , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Fibroblastos/metabolismo , Proliferação de Células , Inflamação/metabolismo , Células CultivadasRESUMO
BACKGROUND: Studies involving human fibroblasts and use of human growth hormone (HGH) administration for injury recovery are limited. It is plausible that if the administration of HGH to human cells increased cellular proliferation and differentiation, then HGH might be able to assist in accelerating recovery from injury. HYPOTHESIS: HGH will increase proliferation and differentiation of human tendon and ligament fibroblasts in vitro based on both a single-dose and a sustained-dose model of HGH administration. STUDY DESIGN: Basic science cellular study. METHODS: Human tendon and ligament tissue were harvested from 24 patients. Tissue samples were digested with type I collagenase to isolate the target cell types. HGH was administered directly to isolated cells at doses ranging from 100 pg/mL to 10 µg/mL, either in a single-dose or a sustained-dose model. Proliferation was analyzed at days 4 and 7. Differentiation of ligament and tendon fibroblasts was assessed at day 14. RESULTS: Administration of a single-dose of HGH to both cell types demonstrated similar or inferior cellular proliferation compared with controls after 7 days. For the sustained-dosing model of ligament fibroblasts, only the 100 ng/mL concentration demonstrated at least statistically similar or improved proliferation compared with controls. When examining the 100 ng/mL HGH concentration with larger sample sizes, cellular proliferation was not improved over controls for any cell type for the single- or sustained-dosing models. Proliferation for tendon fibroblasts was either similar or inferior to the control group at all concentrations of HGH. There was no clear dose-response relationship demonstrating enhanced collagen production with administration of HGH to suggest it enhances injury recovery. CONCLUSION: HGH administered to human tendon and ligament fibroblasts does not appear to positively affect cellular proliferation and differentiation. CLINICAL RELEVANCE: This study does not support the use of HGH for accelerating recovery from injury.
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The psychedelic beverage ayahuasca is originally obtained by Banisteriopsis caapi (B. caapi) (BC) and Psychotria viridis (P. viridis) (PV). However, sometimes these plant species are replaced by others that mimic the original effects, such as Mimosa hostilis (M. hostilis) (MH) and Peganum harmala (P. harmala) (PH). Its worldwide consumption and the number of studies on its potential therapeutic effects has increased. This study aimed to evaluate the anticancer properties of ayahuasca in human colorectal adenocarcinoma cells. Thus, the maximum inhibitory concentration (IC50) of decoctions of MH, PH, and a mixture of these (MHPH) was determined. The activities of caspases 3 and 9 were evaluated, and the cell proliferation index was determined through immunocytochemical analysis (Ki-67). Two fluorescent probes were used to evaluate the production of oxidative stress and the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) was also evaluated. It was demonstrated that exposure to the extracts significantly induced apoptosis in Caco-2 cells, while decreasing cell proliferation. MH and MHPH samples significantly reduced oxidative stress and significantly increased glutathione peroxidase activity. No significant differences were found in SOD activity. Overall, it was demonstrated that the decoctions have a potential anticancer activity in Caco-2 cells.
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WDR68, a conserved WD40 repeat-containing protein, interacts with E1A and is involved in the E1A-induced cell proliferation and oncogenic transformation, but the intrinsic molecular mechanisms of this process remain to be elucidated. Here, we demonstrate that WDR68 promotes the proliferation of 293T cells by interacting with a series of ribosome biogenesis-regulating proteins. Gene Set Enrichment Analysis (GSEA) of RNA-seq data also revealed that the ribosome biogenesis-associated gene signatures could be the most significantly enriched in the WDR68 expression groups. In accordance, 293T cells are more sensitive to the ribosome biogenesis inhibitors than 293 cells. Taken together, our results indicated that WDR68 could promote cell proliferation through the activation of ribosome biogenesis in the 293T cell context. This provides new insights into the understanding of the function of WDR68 and the molecular characterisation of 293T tool cells.
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Proliferação de Células , Ribossomos , Humanos , Ribossomos/metabolismo , Células HEK293 , Ligação Proteica , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genéticaRESUMO
BACKGROUND/AIM: Radiation therapy is pivotal in cancer treatment; however, its efficacy is limited by challenges such as tumor recurrence. This study delves into the role of exosomes, which are molecular cargo-bearing vesicles, in influencing cell proliferation, radioresistance, and consequent post-irradiation tumor recurrence. Given the significance of exosomes from irradiated malignancies in diagnostics and therapy, it is vital to delineate their functional dynamics, especially in breast and cervical cancer cell lines, where the impact of irradiation on exosome behavior remains enigmatic. MATERIALS AND METHODS: Using MDA-MB-231 and HeLa cell lines, exosomes were isolated from the culture supernatant via ultracentrifugation. The bicinchoninic acid assay was used to measure exosome quantities in irradiated and non-irradiated cells. Radiosensitivity was assessed using colony formation assays, while the role of the MAPK/Erk signaling pathway in recipient cell proliferation and radioresistance was probed using western blotting. RESULTS: Irradiated cells, in both MDA-MB-231 and HeLa lines, produced significantly more exosomes than their non-irradiated counterparts. Co-culturing irradiated cells with exosomes led to increased cell survival post-irradiation and enhanced cell proliferation in both cell lines. Western blotting indicated elevated p-Erk expression in such cells, underscoring the influence of the MAPK/Erk pathway in radioresistance and proliferation. CONCLUSION: The study establishes a potential nexus between exosome secretion and tumor resurgence following radiotherapy. The spotlight falls on the MAPK/ERK signaling conduit as a key influencer. This new knowledge provides an innovative strategy for counteracting cancer recurrence after radiotherapy, emphasizing the importance of understanding the multifaceted roles of exosomes in this context.
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Exossomos , Sistema de Sinalização das MAP Quinases , Humanos , Células HeLa , Exossomos/metabolismo , Recidiva Local de Neoplasia/patologia , Proliferação de Células , Linhagem Celular TumoralRESUMO
In this study, electrospun scaffolds were fabricated using polycaprolactone (PCL) loaded with varying concentrations of ß-carotene (1.2%, 2.4%, and 3.6%) via the electrospinning technique. The electrospinning process involved the melting of PCL in acetic acid, followed by the incorporation of ß-carotene powder under constant stirring. Raman spectroscopy revealed a homogeneous distribution of ß-carotene within the PCL matrix. However, the ß-carotene appeared in particulate form, rather than being dissolved and blended with the PCL matrix, a result also confirmed by thermogravimetric analysis. Additionally, X-ray diffraction analysis indicated a decrease in crystallinity with increasing ß-carotene concentration. Mechanical testing of the scaffolds demonstrated an increase in ultimate strain, accompanied by a reduction in ultimate stress, indicating a potential plasticizing effect. Moreover, antimicrobial assays revealed a marginal antibacterial effect against Escherichia coli for scaffolds with higher ß-carotene concentrations. Conversely, preliminary biological assessment using KUSA-A1 mesenchymal cells indicated enhanced cellular proliferation in response to the scaffolds, suggesting the potential biocompatibility and cell-stimulating properties of ß-carotene-loaded PCL scaffolds. Overall, this study provides insights into the fabrication and characterization of electrospun PCL scaffolds containing ß-carotene, laying the groundwork for further exploration in tissue engineering and regenerative medicine applications.
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The MTCH2 protein is located on the mitochondrial outer membrane and regulates mitochondria-related cell death. This study set out to investigate the role of MTCH2 in the underlying pathophysiological mechanisms of breast cancer (BC). MTCH2 expression levels in BC were analyzed using bioinformatics prior to verification by cell lines in vitro. Experiments of over-expression and siRNA-mediated knockdown of MTCH2 were conducted to assess its biological functions, including its effects on cellular proliferation and cycle progression. Xenografts were utilised for in vivo study and signaling pathway alterations were examined to identify the mechanisms driven by MTCH2 in BC proliferation and cell-cycle regulation. MTCH2 was up-regulated in BC and correlated with patients' overall survival. Over-expression of MTCH2 promoted cellular proliferation and cycle progression, while silencing MTCH2 had the opposite effect. Xenograft experiments were utilised to confirm the in vitro cellular findings and it was identified that the PI3K/Akt signaling pathway was activated by MTCH2 over-expression and suppressed by its silencing. Moreover, the activation of IGF-1R rescued cellular growth and cycle arrest induced by MTCH2-silencing. Overall, this study reveals that expression of MTCH2 in BC is upregulated and potentiates cellular proliferation and cycle progression via the PI3K/Akt pathway.
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In recent years, rising temperatures have caused heat stress (HS), which has had a significant impact on livestock production and growth, presenting considerable challenges to the agricultural industry. Research has shown that miR-425-5p regulates cellular proliferation in organisms. However, the specific role of miR-425-5p in bovine mammary epithelial cells (BMECs) remains to be determined. The aim of this study was to investigate the potential of miR-425-5p in alleviating the HS-induced proliferation stagnation in BMECs. The results showed that the expression of miR-425-5p significantly decreased when BMEC were exposed to HS. However, the overexpression of miR-425-5p effectively alleviated the inhibitory effect of HS on BMEC proliferation. Furthermore, RNA sequencing analysis revealed 753 differentially expressed genes (DEGs), comprising 361 upregulated and 392 downregulated genes. Some of these genes were associated with proliferation and thermogenesis through enrichment analyses. Further experimentation revealed that TOB2, which acts as a target gene of miR-425-5p, is involved in the regulatory mechanism of BMEC proliferation. In summary, this study suggests that miR-425-5p can promote the proliferation of BMECs by regulating TOB2. The miR-425-5p/TOB2 axis may represent a potential pathway through which miR-425-5p ameliorates the proliferation stagnation of BMECs induced by HS.
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Glândulas Mamárias Animais , MicroRNAs , Animais , Bovinos , Proliferação de Células/genética , Células Epiteliais/metabolismo , Expressão Gênica , MicroRNAs/metabolismo , Glândulas Mamárias Animais/citologia , FemininoRESUMO
BACKGROUND AND OBJECTIVE: Chronic rhinosinusitis is a common inflammatory disorder in sinonasal mucosa that could be developed with or without nasal polyps. Cellular proliferation is suggested as a possible mechanism of nasal polyp development. However, conducted studies in this context are limited. So, the present study's aim is the comparison of Proliferating cell nuclear antigen (PCNA) expression in nasal polyps and chronic rhinosinusitis. MATERIALS AND METHODS: In this cross-sectional study, 70 nasal polyp and 60 chronic rhinosinusitis samples from patients referred to Mostafa Khomeini Hospital, Tehran from 2017 to 2022 were immunohistochemically stained by PCNA marker. The percentage of PCNA nuclear expression was determined in two groups and its association with the type of pathological lesion and the patient's age and sex was analyzed by SPSS statistic software version 24 statistical software (IBM Statistics, USA). RESULTS: The mean expression of PCNA in nasal polyp and chronic rhinosinusitis samples was 16.55%⯱â¯13.66 and 17.58%⯱â¯12.68 respectively (ranging from 0 to 57% in both groups) however, there was no significant statistical difference between the two groups (pâ¯=â¯0.479). No relationship was found between PCNA expression with age and sex in none of the chronic rhinosinusitis and nasal polyp groups. CONCLUSION: Proliferative activity of the nasal epithelial cell is similar in chronic rhinosinusitis with and without nasal polyps and it is considered that the increase of epithelial cell proliferative activity probably has no role in nasal polyp development in patients with chronic rhinosinusitis.
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The active form of vitamin D, 1,25-dihydroxyvitamin D3, is known to act via VDR (vitamin D receptor), affecting several physiological processes. In addition, PDIA3 (protein disulphide-isomerase A3) has been associated with some of the functions of 1,25-dihydroxyvitamin D3. In the present study we used siRNA-mediated silencing of PDIA3 in osteosarcoma and prostate carcinoma cell lines to examine the role(s) of PDIA3 for 1,25-dihydroxyvitamin D3-dependent responses. PDIA3 silencing affected VDR target genes and significantly altered the 1,25-dihydroxyvitamin D3-dependent induction of CYP24A1, essential for elimination of excess 1,25-dihydroxyvitamin D3. Also, PDIA3 silencing significantly altered migration and proliferation in prostate PC3 cells, independently of 1,25-dihydroxyvitamin D3. 1,25-Dihydroxyvitamin D3 increased thermostability of PDIA3 in cellular thermal shift assay, supporting functional interaction between PDIA3 and 1,25-dihydroxyvitamin D3-dependent pathways. In summary, our data link PDIA3 to 1,25-dihydroxyvitamin D3-mediated signalling, underline and extend its role in proliferation and reveal a novel function in maintenance of 1,25-dihydroxyvitamin D3 levels.
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Movimento Celular , Proliferação de Células , Isomerases de Dissulfetos de Proteínas , Receptores de Calcitriol , Vitamina D3 24-Hidroxilase , Isomerases de Dissulfetos de Proteínas/metabolismo , Isomerases de Dissulfetos de Proteínas/genética , Humanos , Receptores de Calcitriol/metabolismo , Receptores de Calcitriol/genética , Linhagem Celular Tumoral , Vitamina D3 24-Hidroxilase/genética , Vitamina D3 24-Hidroxilase/metabolismo , Calcitriol/farmacologia , Calcitriol/metabolismo , Inativação Gênica , RNA Interferente Pequeno/metabolismo , RNA Interferente Pequeno/genética , Vitamina D/metabolismo , Vitamina D/farmacologia , Vitamina D/análogos & derivados , Masculino , Neoplasias da Próstata/genética , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologiaRESUMO
Introducción: El Ki67 es una proteína reguladora del ciclo celular asociada a la proliferación de las células tumorales. Su expresión siempre ha tenido un papel en la clasificación tumoral, constituye uno de los factores pronósticos y predictivos en el carcinoma mamario. Objetivo: Determinar la relación entre la expresión del marcador de Ki67 y otros factores pronósticos clásicos del cáncer de mama. Métodos: Se realizó un estudio descriptivo analítico, de corte transversal, realizado en el Hospital Clínico-Quirúrgico Docente Celestino Hernández, Villa Clara, entre enero 2017 y mayo de 2019. Se incluyeron 286 mujeres con diagnóstico de carcinoma de mama infiltrante, a cuyas biopsias se les realizó estudio inmunohistoquímico. La expresión del marcador celular Ki67 fue categorizado como baja (Ki6720 %). Se analizó la relación entre el nivel de expresión de Ki67 con otros factores pronósticos y predictivos del carcinoma mamario. Resultados: El tipo histológico no especial (carcinoma ductal) fue el que se reportó con mayor frecuencia. Los niveles de expresión altos del marcador celular Ki67 (Ki67≥20 %) se asociaron con el grado histológico alto (grado 3) y la sobreexpresión de Her2. La expresión baja del Ki-67 (<20 %) se asoció con la expresión de los receptores de estrógeno y progesterona. No se demostró asociación significativa entre la talla tumoral y la expresión de Ki67. Conclusiones: Los niveles de expresión del Ki67 mostraron una asociación significativa con varios factores predictivos y pronósticos clásicos del cáncer de mama.
Introduction: Ki67 is a regulatory protein of cellular cycle which is associated to the proliferation of tumoral cells. Its expression has always had an important role at the tumor classification and it is one of the prognostic and predictive factors in breast carcinoma. Objective: To determine the relationship between the expression of Ki67 and other classic prognostic factors used in breast cancer. Methods: A cross-sectional, analytic and descriptive study was carried out at the Teaching Clinic-Surgical Hospital Celestino Hernández, Villa Clara, from January 2017 to June 2019. It was included 286 women with diagnosis of infiltrating breast carcinoma, whose biopsies were studied by immunohistochemistry. The Ki-67 cell marker expression was categorized as low (Ki-6720 %). It was analyzed the relationship between level of expression of Ki67 and other classical prognostic and predictive factors. Results: The no special histological type (ductal carcinoma) was the type more often reported. High expression level of Ki67 was associated with the high histological grade (grade 3) and the overexpression of Her2. Low expression of Ki-67 (<20 %) was associated with the expression of estrogen and progesterone receptors. There was not significant association between the tumor size and the expression of Ki67. Conclusions: The levels of expression of Ki67 showed significant association with several predictive and prognostic factors of breast carcinoma.
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ABSTRACT Objective To investigate if ICI 182,780 (fulvestrant), a selective estrogen receptor alpha/beta (ERα/ERβ) antagonist, and G-1, a selective G-protein-coupled receptor (GPER) agonist, can potentially induce autophagy in breast cancer cell lines MCF-7 and SKBr3, and how G-1 affects cell viability. Methods Cell viability in MCF-7 and SKBr3 cells was assessed by the MTT assay. To investigate the autophagy flux, MCF-7 cells were transfected with GFP-LC3, a marker of autophagosomes, and analyzed by real-time fluorescence microscopy. MCF-7 and SKBr3 cells were incubated with acridine orange for staining of acidic vesicular organelles and analyzed by flow cytometry as an indicator of autophagy. Results Regarding cell viability in MCF-7 cells, ICI 182,780 and rapamycin, after 48 hours, led to decreased cell proliferation whereas G-1 did not change viability over the same period. The data showed that neither ICI 182,780 nor G-1 led to increased GFP-LC3 puncta in MCF-7 cells over the 4-hour observation period. The cytometry assay showed that ICI 182,780 led to a higher number of acidic vesicular organelles in MCF-7 cells. G-1, in turn, did not have this effect in any of the cell lines. In contrast, ICI 182,780 and G-1 did not decrease cell viability of SKBr3 cells or induce formation of acidic vesicular organelles, which corresponds to the final step of the autophagy process in this cell line. Conclusion The effect of ICI 182,780 on increasing acidic vesicular organelles in estrogen receptor-positive breast cancer cells appears to be associated with its inhibitory effect on estrogen receptors, and GPER does notseem to be involved. Understanding these mechanisms may guide further investigations of these receptors' involvement in cellular processes of breast cancer resistance.
RESUMO Objetivo Avaliar o efeito dos compostos ICI 182,780 (fulvestranto), um antagonista seletivo dos receptores de estrógeno alfa/beta (REα/REβ), e do G-1, um agonista seletivo de receptores de estrógeno acoplados a proteínas-G (GPER), na possível indução de autofagia em linhagens de câncer de mama MCF-7 e SKBr3, bem como o efeito de G-1 na viabilidade celular. Métodos A viabilidade celular de células MCF-7 e SKBr3 foi avaliada pelo ensaio com MTT. Para investigar a indução da autofagia, células MCF-7 foram transfectadas com GFP-LC3, um marcador de autofagossomos, e analisadas por microscopia de fluorescência em tempo real. As células MCF-7 e SKBr3 foram incubadas com o indicador de compartimentos ácidos laranja de acridina e analisadas por citometria de fluxo como indicativo para autofagia. Resultados Em células MCF-7, o ICI 182,780 e rapamicina após 48 horas levaram à diminuição da viabilidade celular, enquanto o G-1 não alterou a viabilidade no mesmo período de tratamento. Nem o ICI 182,780 e nem o G-1 induziram aumento na pontuação de GFP-LC3 em células MCF-7 até 4 horas. Já os ensaios de citometria de fluxo demonstraram que ICI 182,780 levou ao aumento de compartimentos ácidos em células MCF-7. O G-1 não aumentou estes parâmetros em ambas as linhagens. Por outro lado, ICI 182,780 e G-1 não induziram à redução da viabilidade em células SKBr3 e nem à formação de compartimentos ácidos, como etapa final do processo autofágico. Conclusão O aumento de compartimentos ácidos pelo ICI 182,780 em células de câncer de mama positivas para receptores de estrógeno parece estar associado com seu efeito inibidor de receptores de estrógeno, mas sem o envolvimento de GPER. A compreensão desses mecanismos pode direcionar estudos sobre o envolvimento dos receptores nos processos celulares de resistência do câncer de mama.
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Humanos , Feminino , Autofagia/efeitos dos fármacos , Neoplasias da Mama/patologia , Neoplasias da Mama/tratamento farmacológico , Receptores Acoplados a Proteínas G/agonistas , Antagonistas do Receptor de Estrogênio/farmacologia , Fulvestranto/farmacologia , Fatores de Tempo , Transfecção/métodos , Sobrevivência Celular/efeitos dos fármacos , Western Blotting , Reprodutibilidade dos Testes , Análise de Variância , Sirolimo/farmacologia , Receptores Acoplados a Proteínas G/análise , Receptor alfa de Estrogênio/antagonistas & inibidores , Receptor beta de Estrogênio/antagonistas & inibidores , Proliferação de Células/efeitos dos fármacos , Células MCF-7 , Citometria de Fluxo/métodosRESUMO
ABSTRACT Eriosema campestre var. macrophylum (Grear) Fortunato, Fabaceae, is a native plant of the Brazilian Cerrado and the decoction of its roots has been used by folk medicine for the therapy of inflammatory diseases. In this study we aimed to investigate the effect of the dichloromethane–ethanolic extract of E. campestre roots on the proliferative response of lymphocytes and to examine the profile of IL-2 production. The effect of dichloromethane–ethanolic extract of E. campestre on the proliferation of phytohemagglutinin-stimulated lymphocytes was evaluated by using flow cytometry and the cell supernatants were assayed for IL-2 concentrations by using an enzyme-linked immunosorbent assay. The phytochemical screening of E. campestre roots was performed to determine the main secondary metabolites through chromogenic and precipitation reactions and by using HPLC-PAD. In addition to the presence of subclasses of flavonoids (flavones and flavonols) in dichloromethane–ethanolic extract of E. campestre, we observed that the extract induced a concentration-dependent decrease in IL-2 levels on the supernatant of the cell cultures as well as an antiproliferative effect on T lymphocytes, including CD4+ and CD8+ cells. The anti-inflammatory effects attributed to E. campestre by folk medicine may partly be explained by its antiproliferative action on T lymphocytes.
RESUMO
Las estatinas son inhibidores competitivos de la 3-hidroxi-3-metilglutaril-coenzima A (HMG-CoA) reductasa ampliamente usados en los tratamientos contra las hipercolesterolemias. Los monoterpenos son componentes no nutritivos de la dieta presentes en aceites esenciales de varias plantas que han demostrado tener múltiples efectos en la vía del mevalonato. Se estudia el efecto y mecanismo de acción de monoterpenos presentes en aceites esenciales, así como la combinación de éstos entre sí y con simvastatina sobre la síntesis de colesterol, el metabolismo lipídico y la proliferación celular in vitro en células hepáticas Hep G2 y no hepáticas A549, e in vivo en ratones atímicos huéspedes y no huéspedes de tumores derivado de células A549 implantados en ellos. Se abre así una gran expectativa sobre la potencialidad de la administración conjunta de distintos monoterpenos y de extractos naturales de aceites esenciales en el mejoramiento de las terapias antihipercolesterolemiantes y/o el tratamiento del cáncer, como así también en el potencial sinergismo con estatinas como una alternativa para disminuir las dosis efectivas y los efectos indeseados y/o tóxicos.
Statins are competitive inhibitors of HMG-CoA reductase used in hypercholesterolemic patients. Monoterpenes are non-nutritive dietary components found in the essential oils of many plants with pharmacologic effects on mevalonate metabolism. The study is centered on the effects and action mechanisms of the monoterpene components of essential oils and the combination of monoterpenes between them and combined with simvastatin on cholesterogenesis, lipid metabolism and cellular proliferation in vitro using two established cell lines, Hep G2 (derived from a human hepatoblastoma), A549 (derived from a human lung adenocarcinoma) and in vivo in no host and host nude mice carrying implanted tumors derived from A549. This opens up great expectations about the potential of co-administration of different natural isoprenoids and essential oils in improving anti-cholesterolemic therapies and/or cancer treatment as well as in the potential synergism with statins as an alternative to lower effective doses, decreasing the likelihood of undesired and/or toxic effects.
As estatinas são inibidores competitivos da 3-hidroxi-3-metilglutaril - coenzima A (HMG-CoA) reductase amplamente utilizados nos tratamentos contra as hipercolesterolemias. Os monoterpenos são componentes não nutritivos da dieta encontrados em óleos essenciais de várias plantas que demonstraram ter múltiplos efeitos na via do mevalonato. Estudamos o efeito e o mecanismo de ação de monoterpenos encontrados em óleos essenciais, bem como a combinação deles entre si e com sinvastatina sobre a síntese de colesterol, o metabolismo lipídico e a proliferação celular in vitro em células hepáticas Hep G2 e não hepáticas A549 e in vivo em camundongos atímicos hospedeiros ou não hospedeiros de tumores derivados de células A549 implantadas neles. Isto abre grandes expectativas sobre o potencial da co-administração de diferentes monoterpenos e de extratos naturais de óleos essenciais na melhoria das terapias anti-hipercolesterolemiantes e/ou tratamento do câncer, assim como no potencial sinergismo com estatinas como uma alternativa para reduzir as doses efetivas e os efeitos indesejáveis e/ou tóxicos.
Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases/metabolismo , Monoterpenos/metabolismo , Células A549 , Anticolesterolemiantes , Células Hep G2 , HepatócitosRESUMO
El Granuloma Piogénico (GP) es una lesión no neoplásica de la cavidad bucal y de la piel relativamente poco frecuente y extremadamente rara en el tracto gastrointestinal. En la cavidad bucal donde es a menudo encontrada sobre tejido queratinizado. Se considera una hiperplasia inflamatoria. Clínicamente es una lesión uniforme o exofítica lobulada, pequeña, pápulas rojas eritematosas sobre una base pediculada o a veces sésil y que habitualmente sangra. El diámetro varía de pocos milímetros a varios centímetros. El propósito de este artículo es presentar un caso de Granuloma Piogénico, y valorar los factores que intervienen en su patogenia, así como la clínica, diagnóstico diferencial y tratamiento
Pyogenic granuloma (PG) is a non-neoplastic tumor lesion of the bucal cavity and skin relatively rare and extremely rare in the gastrointestinal tract. In the bucal cavity where it is often found on keratinized tissue. It is considered an inflammatory hyperplasia. Clinically it is an uniform or lobulated exophytic lesion, small, red erythematous papules on a pedunculated or sometimes sessile base that usually bleed. The diameter varies from a few millimeters to several centimeters. The purpose of this paper is to present a case of pyogenic granuloma, and evaluate the factors involved in its pathogenesis and the clinical differential diagnosis and treatment