Cyclin D1 expression in penile cancer.

The main goal of the present study was to analyze the expression profile of cyclin D1 in patients with PC, and to determine possible correlations with clinical and histopathological features. A survey was conducted with 100 patients diagnosed with PC, who were treated at two reference hospitals in São Luís, Maranhão, Brazil, between 2013 and 2017. A review of clinical, epidemiological, and histopathological data was performed, Human Papillomavírus (HPV) DNA was detected using polymerase chain reaction (PCR) and cyclin D1 expression analysis was performed using immunohistochemical techniques. The data revealed that the absence of cyclin D1 expression was significantly associated with HPV-positive histological subtypes (p = 0.001), while its expression was associated with high-grade tumors (p = 0.014), histological subtype (p = 0.001), presence of sarcomatoid transformation (p = 0.04), and perineural invasion (p = 0.023). Patients with cyclin D1 expression exhibited lower disease-free survival compared to the cyclin D1-negative group, although the difference was not statistically significant. The results suggest that cyclin D1 may be a potential biomarker for PC, especially for poorer prognosis.

FLNA expression modulates pathological markers of pituitary neuroendocrine tumours.

Due to the current limited knowledge about the role of filamin A (FLNA) in pituitary tumour progression, we aimed to analyse FLNA expression levels and its impact on aggressive markers of pituitary neuroendocrine tumours (PitNETs), using an integrative approach of in vivo and in vitro models and human samples. An increase in the expression levels of FLNA was observed in the advanced tumoural stages of the hyperplastic adenomatous pituitary model, concomitant with a decrease in cell proliferation and with a modification in the subcellular localisation of this protein. Similarly, overexpression of FLNA in the somatolactotropic GH3 cell line induced a decrease in the cell proliferation, promoted a migratory phenotype, increased invasion activity, and decreased the prolactin secretion. Cyclin D1 (CCND1) and cyclin-dependent kinase 4 (CDK4) expression increased in both models in correlation with the increase observed in FLNA levels. When human tissues were analysed a significant increase of FLNA was observed in PitNETs compared to normal pituitary gland, with heterogeneous intracellular localisation. Higher levels of FLNA expression were observed in tumours with invasive characteristics. These results underline the crucial roles of FLNA as a modulator of pathological markers and as a potential prognostic marker in pituitary tumours.

Synergistic effect of chrysin and radiotherapy against triple-negative breast cancer (TNBC) cell lines.

PURPOSE: Triple-negative breast cancer (TNBC) is the most aggressive form of breast cancer, accounting for 20% of cases. Due to the lack of a molecular target, limited options are available for TNBC treatment. Radiation therapy (RT) is a treatment modality for the management of TNBC following surgery; however, it has a detrimental effect on surrounding healthy tissues/cells at a higher rate. METHODS: We examined the effect of RT in combination with chrysin as a possible radiosensitizing agent in an MDA-MB-231 cell line as a model of a TNBC. The growth inhibitory effects of chrysin were examined using an MTT assay. Flow cytometry was performed to evaluate apoptosis and expression of hypoxia-induced factor-1α (HIF-1α). The protein expression of p-STAT3/STAT3 and Cyclin D1 was examined using western blotting. Real-time PCR determined apoptotic-related genes (Bax, BCL2, p53). RESULTS: Treatment of MDA-MB-231 cells with chrysin in combination with RT caused synergistic antitumor effects, with an optimum combination index (CI) of 0.495. Our results indicated that chrysin synergistically potentiated RT-induced apoptosis in MDA-MB-231 compared with monotherapies (chrysin and/or RT alone). Expression of HIF-1α was decreased in the cells exposed to combinational therapy. The apoptotic effect of combinational therapy was correlated with increased Bax (pro-apoptotic gene) and p53 levels along with reduced expression of Bcl-2 (anti-apoptotic gene). Increased apoptosis was associated with reduced expression of Cyclin D1, p-STAT3. CONCLUSION: These findings highlight the potential effect of chrysin as a radiosensitizer, indicating the synergistic anti-cancer effect of chrysin and RT in TNBC. Further investigation is warranted in this regard.

Paraganglioma del cuerpo carotídeo asociado a un adenoma paratiroideo: reporte de caso / Carotid body paraganglioma associated with a parathyroid adenoma: case report and literature review

Introducción: Los paragangliomas son tumores raros, y más raros aun cuando se presentan con otros tumores endocrinos. Objetivo: Presentar el caso de un paraganglioma del cuerpo carotídeo asociado con adenomas paratiroideos. Materiales y Método: Se presenta la evaluación clínica, imagenológica y fotográfica del caso. Resultados: Se presenta el caso de una paciente de 34 años con una masa cervical en región cervical de un año de evolución y que, durante los estudios de extensión, se encontró que correspondía a un paraganglioma en la bifurcación carotídea izquierda, asociada además con dos adenomas paratiroideos, que fueron resecados sin complicaciones. Discusión: Se discute la fisiopatología, el diagnóstico y manejo en relación con el caso presentado. Conclusión: La presentación de paragangliomas del cuerpo carotídeo asociadas con adenomas paratiroideos es rara, y su evaluación clínica deberá ser individualizada, dado que, si bien el manejo será en su mayoría quirúrgico, el abordaje dependerá de cada caso.

Introduction: Paragangliomas are rare tumors, and they are even rarer when they present with other endocrine tumors. Aim: To present a clinical case of a carotid body paraganglioma associated with parathyroid adenomas. Materials and Method: There are shown the clinical evaluation, images, and photos of the case. Results: We present the case of a 34 years old female patient with a cervical mass, which has grown for a year, and, after extension studies, it was found that the mass corresponded to a paraganglioma located in the left carotid bifurcation, and it was associated with two parathyroid adenomas, all the neoplasms were resected with no complications. Discussion: It is discussed physiopathology, diagnosis and management based on the presented case Conclusion: Carotid body paragangliomas associated with parathyroid adenomas are rare, and the clinical evaluation must be individual, given that, most of the management is surgical, however, the approach will depend on each case.

Biomarkers involved in the proliferation of the odontogenic keratocyst, glandular odontogenic cyst and botryoid odontogenic cyst.

INTRODUCTION: Odontogenic cysts are a heterogeneous group of lesions with varied clinical behavior. OBJECTIVE: To analyze the expression of the epidermal growth factor receptor (EGFR), Cyclin D1, and transcription factor SOX2 in the odontogenic epithelium evaluating the cell cycle control and cystic expansion. METHODS: This was a cross-sectional study including 40 cases, 20 odontogenic keratocysts (OKC), 10 botryoid odontogenic cysts (BOC), and 10 glandular odontogenic cysts (GOC). RESULTS: All cases of OKC, BOC, and GOC were positive for EGFR in all layers of the cyst lining. The highest expression of nuclear Cyclin D1 was observed in the suprabasal layer of OKCs and in the basal and suprabasal layers of GOC and BOC (p < 0.001). In addition, SOX2 was only expressed in the suprabasal layer of OKCs. CONCLUSION: The high expression of EGFR in the cyst membrane suggests that EGF stimulates epithelial proliferation in BOCs, and the high expression of SOX2 in OKCs may be related to the presence of stem cells in the lesion. Cyclin D1 is related to cell cycle disruption in G1-S contributing to stimulates epithelial proliferation of OKCs and GOCs and BOCs.

p53 & Cyclin D1 expression in surgically resected clear margins of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of the most well-known malignancies that affect the human population worldwide. The early diagnosis and early intervention of OSCC help improve the survival rate of the patients. The tumour free surgical margins are a positive prognostic factor for recurrence-free survival. The molecular markers can be used to detect the tumour free surgical margins. Aim: The aim of the study is to evaluate the expression of p53 & Cyclin D1 marker in resected surgical apparently clear margins and to correlate the p53 & Cyclin D1 expression with clinicopathological characteristics and patient outcome. Methods: The study population included retrospective cases of OSCC with apparently clear margins (2017-18) n=10 and Clinicopathological variables relevant to survival analysis were recorded. Finally, two margins were selected from each case, a total of 20 margins were included in this study. Paraffin-embedded wax blocks retrieved and tissue sections were made. Expression of cyclin D1 and p 53 was assessed by the immunohistochemical staining procedure Results: Positive expressions Cyclin D1 in 40% of mild dysplasia margins and 60% in clearance adequate margins were present. p53 expression was seen in 16% of mild dysplasia margins and 84% in clearance adequate margins. The expression of p53 and Cyclin D1 molecular markers are noted in the basal & parabasal layer of epithelium. Conclusion: Molecular markers could play a more reliable method for the assessment of dysplasia at the margins

Oncogenic and tumor suppressor pathways in subchronic aflatoxicosis in rats: Association with serum and urinary aflatoxin exposure biomarkers.

In this study, the changes in oncogenic and tumor suppressor signaling pathways in liver and their association with serum and urinary biomarkers of aflatoxin exposure were evaluated in Wistar rats fed diets containing aflatoxin B1 (AFB1) for 90 days. Rats were divided into four groups (n = 15 per group) and assigned to dietary treatments containing 0 (control), 50 (AFB50), 100 (AFB100) and 200 µg AFB1 kg-1 diet (AFB200). Multiple preneoplastic foci of hepatocytes marked with glutathione-S-transferase-placental form (GST-P) were identified in AFB100 and AFB200 groups. Hepatocellular damage induced by AFB1 resulted in overexpression of cyclin D1 and ß-catenin. The liver expression of retinoblastoma (Rb) and p27Kip1 decreased in AFB100 and AFB200 groups, confirming the favorable conditions for neoplastic progression to hepatocellular carcinoma. All samples from rats fed AFB1-contaminated diets had quantifiable AFB1-lysine in serum or urinary AFM1 and AFB1-N7-guanine, with mean levels of 20.42-50.34 ng mL-1, 5.31-37.68 and 39.15-126.37 ng mg-1 creatinine, respectively. Positive correlations were found between AFB1-lysine, AFM1 or AFB1-N7-guanine and GST-P+, ß-catenin+ and cyclin D1+ hepatocytes, while Rb + cells negatively correlated with those AFB1 exposure biomarkers. The pathways evaluated are critical molecular mechanisms of AFB1-induced hepatocarcinogenesis in rats.

Acute and chronic lithium treatment increases Wnt/ß-catenin transcripts in cortical and hippocampal tissue at therapeutic concentrations in mice.

Lithium activates Wnt/ß-catenin signaling leading to stabilization of free cytosolic ß-catenin. The aim of the present study is to evaluate the in vivo effect of acute and chronic lithium treatment on the expression of ß-catenin target genes, addressing its transcripts HIG2, Bcl-xL, Cyclin D1, c-myc, in cortical and hippocampal tissue from adult mice. Lithium doses were established to yield therapeutic working concentrations. In acute treatment, mice received a 300µL of a 350 mg/kg solution of LiCl by gavage, and were euthanized after 2 h, 6 h and 12 h. To determine the effect of chronic treatment, animals were continuously fed either with chow supplemented with 2 g/kg Li2CO3, or regular chow (controls), being euthanized after 30 days. All animals had access to drinking water and 0.9% saline ad libitum. After acute and chronic treatments samples of peripheral blood were obtained from the tail vein for each animal, and serum concentrations of lithium were determined. All transcripts were up-regulated in cortical and hippocampal tissues of lithium-treated mice, both under acute and chronic treatments. There was a positive correlation between serum lithium concentrations and the increment in the expression of all transcripts. This effect was observed in all time points of the acute treatment (i.e., 2, 6 and 12 hours) and also after 30 days. We conclude that Wnt/ß-catenin transcriptional response (HIG2, Bcl-xL, Cyclin D1 and c-myc) is up-regulated in the mouse brain in response to acute and chronic lithium treatment at therapeutic concentrations.

A tetraprenylated benzophenone 7-epiclusianone induces cell cycle arrest at G1/S transition by modulating critical regulators of cell cycle in breast cancer cell lines.

Breast cancer is a complex disease and encompassing different types of tumor. Although advances in understanding of the molecular bases of breast cancer biology, the therapeutic proposals available still are not effective. In this scenario, the present study aimed to evaluate the mechanisms associated to antitumor activity of 7-Epiclusianone (7-Epi), a tetraprenylated benzophenone, on luminal A (MCF-7) and claudin-low (Hs 578T) breast cancer cell lines. We found that 7-Epi efficiently inhibited cell proliferation and migration of these cells; however MCF-7 was slightly more responsive than Hs 578T. Cell cycle analysis showed accumulation of cells at G0/G1 phase with drastic reduction of S population in treated cultures. This effect was associated to downregulation of CDKN1A (p21) and cyclin E in both cell lines. In addition, 7-Epi reduced cyclin D1 and p-ERK expression levels in MCF-7 cell line. Cytotoxic effect of 7-Epi on breast cancer cell lines was associated to its ability to increase BAX/BCL-2 ratio. In conclusion, our findings showed that 7-Epi is a promising antitumor agent against breast cancer by modulating critical regulators of the cell cycle and apoptosis.

DNA repair and cell synthesis proteins: immunohistochemical expression and correlation with recurrence-regrowth in meningiomas.

Meningiomas are considered the second most common neoplasm of the central nervous system in adults. Most of them are benign with slow growth, frequent in women and with a high recurrence rate. In tumors, DNA error repair processes lose efficacy, providing mutagenesis and genomic instability. This work evaluated the expression of proteins involved in cell synthesis (cyclin D1) and DNA errors repair (MUTYH, XPF, XPG) in meningiomas, relating them to clinical, tumor and survival variables. The study included 85 patients, with a mean age of 52 ± 13.3 years and most of them women (2:1 ratio). Sixty-seven cases were grade I (79%). Grade II tumors were independent predictors of recurrence-regrowth (HR: 2.8; p = 0.038). The high expression of cyclin D1 was associated with grade II (p = 0.001) and low MUTYH expression with grade I (p = 0.04). Strong expression of XPF and XPG was associated with grade II (p = 0.002; p < 0.001) and with recurrence-regrowth (p = 0.04; p = 0.003). Strong XPF expression was significantly related to large tumors (p = 0.03). An association of cyclin D1, MUTYH and XPF were found. Survival was not associated with the expression of any of the proteins studied. To know the role of DNA repair proteins and cell synthesis is important for understanding the processes of origin and tumor development. Grade II meningiomas and strong expression of XPF and XPG were predictors of recurrence or regrowth and may assist in clinical management, considering the high recurrence of meningiomas and the absence of consensus regarding treatment.

Avaliação morfológica e imuno-histoquímica do potencial proliferativo epitelial das lesões císticas odontogênicas agressivas / Morphological and immunohistochemical evaluation of the epithelial proliferative potential of aggressive odontogenic cystic lesions

INTRODUÇÃO: Os cistos odontogênicos são conhecidos pelo seu potencial de crescimento e reabsorção óssea dos ossos gnáticos, que variam de indolentes a agressivas, independente de sua histogênese, vinculada aos remanescentes epiteliais, ectomesenquimais e/ou mesenquimais da odontogênese. Estas lesões em sua maioria, apresentam patogênese controversa, motivo pelo qual estimulam numerosas investigações sobre a possível relação entre o epitélio odontogênico e o desenvolvimento destas lesões. OBJETIVO: Analisar os diferentes padrões de proliferação do epitélio odontogênico das lesões císticas odontogênicas de comportamento mais agressivo, através da analise da imunoexpessão do fator de crescimento epidérmico (EGFR) e da via de transcrição SOX 2 no controle das fases do ciclo celular (ciclina D1) em lesões odontogênicas de caráter agressivo. MATERIAIS E MÉTODOS: Estudo seccional com amostra não probabilística por conveniência composta por 31 casos, sendo 10 casos de ceratocistos odontogênicos recidivantes, 10 casos de ceratocistos odontogênicos (COs) isolados, 7 casos de cistos odontogênicos botrióides (COB) e 4 casos de cistos odontogênicos glandulares (COG). RESULTADOS: Ao analisar estatisticamente com os testes de Mann-Whitney e Kruskal-Wallis; P <0,05 foi considerado significativo. A maior expressão de células positivas para ciclina D1 foi observada na camada suprabasal de ceratocistos odontogênicos (P <0,001) e nas camadas basal e suprabasal dos COGs (P <0,001), e porções dos COBs (P>0,001). Não houve diferença estatística entre os COs recorrente e não recorrentes. Além disso, todos os casos de COs, COBs e COGs foram positivos para EGFR em todas as camadas do revestimento epitelial cístico. Os COBs e COGs apresentaram negatividade para o SOX 2 enquanto que nos COs foram encontrados alta expressão de SOX 2 na camada suprabasal. CONCLUSÕES: A ciclina D1 implica nos distúrbios do ciclo celular nas fases G1-S refletindo na agressividade dos COs e COGs. A superexpressão do EGFR demonstrada nos COs, COBs e COGs sugere que a combinação citoplasmática e citoplasmática membranosa é indicativo do potencial de proliferação do revestimento epitelial. O alto padrão da expressão do SOX 2 no CO também pode explicar a natureza agressiva da lesão e são responsáveis pela presença de numerosos cistos-filhos responsáveis por sua alta taxa de recorrência. A subpopulação de CSC se apresenta como motivo de recorrência aos COs, pela propriedade de auto-renovação e pluripotência destas células (AU).

INTRODUCTION: Odontogenic cysts are known for their growth potential and bone resorption of gnathic bones, ranging from indolent to aggressive, regardless of their histogenesis, linked to the epithelial, ectomesenchymal and / or mesenchymal remnants of odontogenesis. Most of these lesions have controversial pathogenesis, which is why they stimulate numerous investigations on the possible relationship between the odontogenic epithelium and the development of these lesions. OBJECTIVE: To analyze the different patterns of proliferation of odontogenic epithelium in odontogenic cystic lesions with more aggressive behavior, through the analysis of the epidermal growth factor (EGFR) immunoexpression and the SOX 2 transcription pathway in the control of the cell cycle phases (cyclin D1 ) in aggressive odontogenic lesions. MATERIALS AND METHODS: Sectional study with a non-probabilistic sample for convenience consisting of 31 cases, 10 cases of recurrent odontogenic keratocysts (COs) isolated, 10 cases of botryoid odontogenic cysts (BOC) and 4 cases of odontogenic cysts glandular (COG). RESULTS: When analyzing statistically with the Mann-Whitney and Kruskal-Wallis tests; P <0.05 was considered significant. The highest expression of positive cells for cyclin D1 was observed in the suprabasal layer of odontogenic keratocysts (P <0.001) and in the basal and suprabasal layers of COGs (P <0.001), and portions of COBs (P> 0.001). There was no statistical difference between recurrent and non-recurring COs. In addition, all cases of COs, COBs and COGs were positive for EGFR in all layers of the cystic epithelial lining. COBs and COGs showed negativity for SOX 2 while COs found high expression of SOX 2 in the suprabasal layer. CONCLUSIONS: Cyclin D1 implies cell cycle disorders in G1-S phases, reflecting the aggressiveness of COs and COGs. The overexpression of EGFR demonstrated in COs, COBs and COGs suggests that the combination of cytoplasmic and membranous cytoplasmic is indicative of the proliferation potential of the epithelial lining. The high standard of SOX 2 expression in CO can also explain the aggressive nature of the lesion and are responsible for the presence of numerous child cysts responsible for its high rate of recurrence. The CSC subpopulation presents itself as a reason for recurrence to COs, due to the self-renewal and pluripotency properties of these cells (AU).

Salvia lachnostachys Benth has antitumor and chemopreventive effects against solid Ehrlich carcinoma.

Salvia lachnostachys is an herbaceous plant with anti-inflammatory, analgesic and cytotoxic properties. This study investigated the antitumor effect of an ethanolic extract of Salvia lachnostachys leaves (EES) in a solid Ehrlich carcinoma model. Ehrlich cells were inoculated subcutaneously in the right pelvic member (2 × 106 cells) in female Swiss mice. The animals were treated with vehicle (10 mL kg-1, p.o.), EES (30 and 100 mg kg-1, p.o.), or methotrexate (2.5 mg kg-1, i.p.) for 21 days (early treatment) or 14 days (late treatment) after tumor inoculation, or 10 days before tumor inoculation and continued for 21 days after tumor inoculation (chemopreventive treatment). The acute toxicity test was performed according OECD guidelines Late treatment with EES had no antitumor effect. Early treatment with 100 mg kg-1 EES prevented tumor development, increased tumor necrosis factor-α (TNF-α) levels and decreased tumor superoxide dismutase (SOD) activity, interleukin-10 (IL-10) levels and Cyclin D1 expression, and tumor cell necrosis was observed. Chemopreventive treatment with EES for 10 and 31 days prevented tumor development in the same manner. EES treatment for 31 days decreased hepatic and tumor SOD activity, tumor IL-10 levels and Cyclin D1 expression, and increased tumor reduced glutathione, N-acetylglucosaminidase, reactive oxygen species, lipid peroxidation, TNF-α levels and Nrf2 expression. No toxicity was observed in the acute toxicity assay. In conclusion, EES had an antitumor effect by inhibiting Cyclin D1 expression and increasing inflammation with early and chemopreventive treatment. Modulation of the antioxidant system also contribute for the antitumor effects of EES.

Análise comparativa entre a coloração do azul de toluidina e a imunoexpressão da ciclina D1 em leucoplasias da mucosa bucal com diferentes graus de atipia / Comparative analysis between the coloring of toluidine blue and the immunoexpression of cyclin D1 in leucoplakias of the bucal mucosa with different degrees of atipia

O câncer de boca ocupa uma posição de destaque em relação ao número total de casos registrados no Brasil. A displasia epitelial (DE) oral é um achado histopatológico associado a um risco aumentado de transformação maligna do epitélio oral. A falha nos mecanismos de sinalização celular, no controle do ciclo celular ou nos mecanismos para reparar danos celulares pode favorecer a processos que culminam com a progressão para o câncer. Objetivo: avaliar comparativamente a resposta clínica da marcação do azul de toluidina (AT) e a imunoexpressão da proteína ciclina D1, uma proteína nuclear de grande importância como regulador da transição da fase G1 para fase S do ciclo celular, em leucoplasias oral (LO). Metodologia: avaliamos 12 pacientes que apresentavam, na cavidade bucal, lesões com diagnóstico clínico de LO. O estudo se desenvolveu em duas etapas: clínica e laboratorial. Foi feita marcação com AT e avaliação Imuno-histoquímica, respectivamente. Após análise quantitativa das lâminas, os dados obtidos foram analisados pelo programa BIOESTAT 2.0, por meio dos testes de Spearman e pelo teste de correlações múltiplas de Pearson. Resultados: não foi observada relação entre a marcação clínica do AT, o grau de displasia da lesão e a imunoexpressão da Ciclina D1 em LO. Conclusão: 82% das lesões apresentaram DE em graus variados, confirmando a necessidade de se realizar o diagnóstico histopatológico das LO e o acompanhamento clínico posterior dos pacientes.

Mouth cancer occupies a prominent position in relation to the total number of cases registered in Brazil. Oral epithelial dysplasia (ED) is a histopathological finding associated with an increased risk of malignant transformation of oral epithelium. The failure in the mechanisms of cell signaling, cell cycle control mechanisms or to repair cell damage can favor processes that culminate with the progression to cancer. Objective: evaluate comparatively the clinical response of the toluidine blue marking (TB) and immuno-expression of cyclin D1 protein, a nuclear protein of great importance as a factor regulating the transition from G1 phase to S phase of the cell cycle, in oral leukoplakia (OL). Methodology: we evaluated 12 patients who presented lesions in the oral cavity with clinical diagnosis of OL. The study was developed in two stages: Clinical and Laboratorial. Marking was made with TB and immunohistochemical evaluation, respectively. After quantitative analysis of blades, the data obtained were analyzed by BIOESTAT program 2.0 through Spearman tests and by Pearson multiple correlation test. Results: no relation was observed between clinical marking of TB, the degree of dysplasia of the lesion and the immuno-expression of Cyclin D1 in OL. Conclusion: 82% of the lesions presented DE in varying degrees, confirming the need to perform histopathological diagnosis of the OL and the subsequent clinical monitoring of the patients.

Immunoexpression of cell cycle regulators in canine prostate with proliferative lesions / Imunoexpressão de reguladores do ciclo celular em próstatas caninas com alterações proliferativas

Immunostaining of p21, p27, p53, cyclin D1, c-myc was evaluated in normal canine prostate and prostate with proliferative disorders to verify the interaction between these regulators of cell cycle progression. From 106 samples of canine prostate obtained from a TMA block, 15 were considered normal, 16 diagnosed as benign prostatic hyperplasia (BPH), 30 as proliferative inflammatory atrophy (PIA), 20 as prostatic intraepithelial neoplasia (PIN), and 25 as prostatic carcinoma (PC). There was positive correlation between p21 and p27 for number of stained cells and staining intensity in all conditions and between c-myc and p53 in prostates with PIN. Considering the number of labeled cells, there was positive correlation between p21 and p53 in the normal prostate. Relative to the intensity of staining, there was positive correlation between p21 and p53 in prostate tissue with PIN and between p27 and c-myc in prostates with PIA. A negative correlation between c-myc and cyclin D1 was also identified in the glands with PIN, considering the number of labeled cells, and between p27 and c-myc in the prostates with PC for staining intensity. In conclusion, the expression of p21, p27, p53, cyclin D1 and c-myc varies according to type of proliferative lesion in canine prostate. Taken together, the results indicate low growth potential of the canine PC in the presence of p21 and p27 overexpression, cyclin D1 low expression and regular expression of c-myc, even with the expression of p53 mutant type. Further, it was possible reaffirm the premalignant potential of PIA and PIN in canine prostate.(AU)

A imunomarcação de p21, p27, p53, ciclina D1 e c-myc foi avaliada na próstata canina normal e com desordens proliferativas para verificar quanto a interação desses reguladores na progressão do ciclo celular. Um total de 106 amostras de próstata canina foi obtido a partir de um bloco de TMA, sendo 15 normais, 16 hiperplasia prostática benigna (HPB), 30 atrofia inflamatória proliferativa (PIA), 20 neoplasia intraepitelial prostática (PIN), e 25 carcinoma prostático (PC). Foi encontrada diferença na imunomarcação de p21, p27, ciclina D1 e p53 no epitélio acinar em relação aos diagnósticos. Houve correlação positiva entre p21 e p27 para as variáveis número de células marcadas e intensidade de imunomarcação em todos os diagnósticos (normal, HPB, PIA, PIN e PC), e entre c-myc e p53 nas próstatas com PIN. De acordo com o número de células marcadas, houve correlação positiva entre p21 e p53 na próstata normal. De acordo com a intensidade de imunomarcação houve correlação positiva entre p21 e p53 no tecido prostático com PIN e entre p27 e c-myc em próstatas com PIA. Foi observada correlação negativa entre c-myc e ciclina D1 nas glândulas com PIN, considerando o número de células marcadas, e entre p27 e c-myc na próstata com PC, para a variável intensidade de imunomarcação. Conclui-se que a expressão de p21, p27, p53, ciclina D1 e c-myc varia na próstata canina de acordo com o tipo de lesão proliferativa. Em conjunto, os resultados indicam baixo potencial de crescimento dos carcinomas da próstata canina quando há superexpressão de p21 e de p27, baixa expressão de ciclina D1 e expressão normal de c-myc, mesmo com expressão de p53 tipo mutante. Ainda, considerando o imunofenótipo semelhante nas glândulas com PIA, PIN e PC no que se refere aos reguladores da progressão do ciclo celular, reitera-se o potencial pré-maligno da PIA e PIN na próstata canina.(AU)

SALL2 represses cyclins D1 and E1 expression and restrains G1/S cell cycle transition and cancer-related phenotypes.

SALL2 is a poorly characterized transcription factor that belongs to the Spalt-like family involved in development. Mutations on SALL2 have been associated with ocular coloboma and cancer. In cancers, SALL2 is deregulated and is proposed as a tumor suppressor in ovarian cancer. SALL2 has been implicated in stemness, cell death, proliferation, and quiescence. However, mechanisms underlying roles of SALL2 related to cancer remain largely unknown. Here, we investigated the role of SALL2 in cell proliferation using mouse embryo fibroblasts (MEFs) derived from Sall2-/- mice. Compared to Sall2+/+ MEFs, Sall2-/- MEFs exhibit enhanced cell proliferation and faster postmitotic progression through G1 and S phases. Accordingly, Sall2-/- MEFs exhibit higher mRNA and protein levels of cyclins D1 and E1. Chromatin immunoprecipitation and promoter reporter assays showed that SALL2 binds and represses CCND1 and CCNE1 promoters, identifying a novel mechanism by which SALL2 may control cell cycle. In addition, the analysis of tissues from Sall2+/+ and Sall2-/- mice confirmed the inverse correlation between expression of SALL2 and G1-S cyclins. Consistent with an antiproliferative function of SALL2, immortalized Sall2-/- MEFs showed enhanced growth rate, foci formation, and anchorage-independent growth, confirming tumor suppressor properties for SALL2. Finally, cancer data analyses show negative correlations between SALL2 and G1-S cyclins' mRNA levels in several cancers. Altogether, our results demonstrated that SALL2 is a negative regulator of cell proliferation, an effect mediated in part by repression of G1-S cyclins' expression. Our results have implications for the understanding and significance of SALL2 role under physiological and pathological conditions.

Increased levels of cyclin D1 negatively impacts on acute lymphoblastic leukemia overall survival

Background: Cyclin D1 is a protein essential for transition from G1 to S phase during cell cycle progression, which has an oncogenic potential and is highly expressed in several human malignancies. However, in view of the heterogeneity of the findings in the literature, the prognostic value of cyclin D1 expression still needs to be validated in different cohorts of adult acute lymphoblastic leukemia (ALL) patients. Methods: Bone marrow samples from 13 healthy donors and 45 adult patients with acute lymphoblastic leukemia were included. Cyclin D1 gene expression was evaluated by quantitative PCR. For statistical analysis, Mann­Whitney test, Fisher's exact test, Chi-squared test and Cox regression were used, as appropriate. All p values were two-sided with a significance level of 5%. Results: Cyclin D1 mRNA levels were similar between primary cells from ALL patients and healthy donors. In ALL patients, high cyclin D1 expression was associated with older age at the diagnosis, presence of BCR-ABL1, and lower white blood cell counts. Importantly, increased cyclin D1 expression was an independent factor that predicted worse overall survival in our adult ALL cohort. Conclusion: Increased levels of cyclin D1 negatively impacted on ALL survival outcome, suggesting that this gene is involved in the malignant phenotype of ALL.

Immunoexpression of cell cycle regulators in canine prostate with proliferative lesions / Imunoexpressão de reguladores do ciclo celular em próstatas caninas com alterações proliferativas

Immunostaining of p21, p27, p53, cyclin D1, c-myc was evaluated in normal canine prostate and prostate with proliferative disorders to verify the interaction between these regulators of cell cycle progression. From 106 samples of canine prostate obtained from a TMA block, 15 were considered normal, 16 diagnosed as benign prostatic hyperplasia (BPH), 30 as proliferative inflammatory atrophy (PIA), 20 as prostatic intraepithelial neoplasia (PIN), and 25 as prostatic carcinoma (PC). There was positive correlation between p21 and p27 for number of stained cells and staining intensity in all conditions and between c-myc and p53 in prostates with PIN. Considering the number of labeled cells, there was positive correlation between p21 and p53 in the normal prostate. Relative to the intensity of staining, there was positive correlation between p21 and p53 in prostate tissue with PIN and between p27 and c-myc in prostates with PIA. A negative correlation between c-myc and cyclin D1 was also identified in the glands with PIN, considering the number of labeled cells, and between p27 and c-myc in the prostates with PC for staining intensity. In conclusion, the expression of p21, p27, p53, cyclin D1 and c-myc varies according to type of proliferative lesion in canine prostate. Taken together, the results indicate low growth potential of the canine PC in the presence of p21 and p27 overexpression, cyclin D1 low expression and regular expression of c-myc, even with the expression of p53 mutant type. Further, it was possible reaffirm the premalignant potential of PIA and PIN in canine prostate.

A imunomarcação de p21, p27, p53, ciclina D1 e c-myc foi avaliada na próstata canina normal e com desordens proliferativas para verificar quanto a interação desses reguladores na progressão do ciclo celular. Um total de 106 amostras de próstata canina foi obtido a partir de um bloco de TMA, sendo 15 normais, 16 hiperplasia prostática benigna (HPB), 30 atrofia inflamatória proliferativa (PIA), 20 neoplasia intraepitelial prostática (PIN), e 25 carcinoma prostático (PC). Foi encontrada diferença na imunomarcação de p21, p27, ciclina D1 e p53 no epitélio acinar em relação aos diagnósticos. Houve correlação positiva entre p21 e p27 para as variáveis número de células marcadas e intensidade de imunomarcação em todos os diagnósticos (normal, HPB, PIA, PIN e PC), e entre c-myc e p53 nas próstatas com PIN. De acordo com o número de células marcadas, houve correlação positiva entre p21 e p53 na próstata normal. De acordo com a intensidade de imunomarcação houve correlação positiva entre p21 e p53 no tecido prostático com PIN e entre p27 e c-myc em próstatas com PIA. Foi observada correlação negativa entre c-myc e ciclina D1 nas glândulas com PIN, considerando o número de células marcadas, e entre p27 e c-myc na próstata com PC, para a variável intensidade de imunomarcação. Conclui-se que a expressão de p21, p27, p53, ciclina D1 e c-myc varia na próstata canina de acordo com o tipo de lesão proliferativa. Em conjunto, os resultados indicam baixo potencial de crescimento dos carcinomas da próstata canina quando há superexpressão de p21 e de p27, baixa expressão de ciclina D1 e expressão normal de c-myc, mesmo com expressão de p53 tipo mutante. Ainda, considerando o imunofenótipo semelhante nas glândulas com PIA, PIN e PC no que se refere aos reguladores da progressão do ciclo celular, reitera-se o potencial pré-maligno da PIA e PIN na próstata canina.

Helicobacter pylori Induced Phosphatidylinositol-3-OH Kinase/mTOR Activation Increases Hypoxia Inducible Factor-1α to Promote Loss of Cyclin D1 and G0/G1 Cell Cycle Arrest in Human Gastric Cells.

Helicobacter pylori (H. pylori) is a human gastric pathogen that has been linked to the development of several gastric pathologies, such as gastritis, peptic ulcer, and gastric cancer. In the gastric epithelium, the bacterium modifies many signaling pathways, resulting in contradictory responses that favor both proliferation and apoptosis. Consistent with such observations, H. pylori activates routes associated with cell cycle progression and cell cycle arrest. H. pylori infection also induces the hypoxia-induced factor HIF-1α, a transcription factor known to promote expression of genes that permit metabolic adaptation to the hypoxic environment in tumors and angiogenesis. Recently, however, also roles for HIF-1α in the repair of damaged DNA and inhibition of gene expression were described. Here, we investigated signaling pathways induced by H. pylori in gastric cells that favor HIF-1α expression and the consequences thereof in infected cells. Our results revealed that H. pylori promoted PI3K/mTOR-dependent HIF-1α induction, HIF-1α translocation to the nucleus, and activity as a transcription factor as evidenced using a reporter assay. Surprisingly, however, transcription of known HIF-1α effector genes evaluated by qPCR analysis, revealed either no change (LDHA and GAPDH), statistically insignificant increases SLC2A1 (GLUT-1) or greatly enhance transcription (VEGFA), but in an HIF-1α-independent manner, as quantified by PCR analysis in cells with shRNA-mediated silencing of HIF-1α. Instead, HIF-1α knockdown facilitated G1/S progression and increased Cyclin D1 protein half-life, via a post-translational pathway. Taken together, these findings link H. pylori-induced PI3K-mTOR activation to HIF-1α induced G0/G1 cell cycle arrest by a Cyclin D1-dependent mechanism. Thus, HIF-1α is identified here as a mediator between survival and cell cycle arrest signaling activated by H. pylori infection.

Casearin D inhibits ERK phosphorylation and induces downregulation of cyclin D1 in HepG2 cells.

Cancer is a public health problem which represents the second cause of death in the world. In this framework, it is necessary to identify novel compounds with antineoplastic potential. Plants are an important source for discovering novel compounds with pharmacological potential. In this study, we aimed to investigate the antiproliferative potential of isolated compounds from Casearia sylvestris on tumor cell lines. Crude extract effectively reduced cell viability of 4 tumor cell lines (HepG2, A549, U251-MG, and HT-144) after 48h treatment. HepG2 and HT-144 were the most responsive cells. Three fractions (aqueous ethanol, n-hexane and ethyl acetate) were tested against HepG2 and HT-144 cells and we observed that compounds with antiproliferative activity were concentrated in n-hexane and ethyl acetate fractions. The casearins A, G and J were isolated from n-hexane fraction, while casearin D was obtained from ethyl acetate fraction. We demonstrated that casearin D significantly inhibited the clonogenic capacity of HepG2 cells after 24h exposure indicating its antiproliferative activity. In addition, G1/S transition cell cycle arrest in HepG2 cells was also observed. These effects are related, at least in part, to ability of the casearin D in reducing ERK phosphorylation and cyclin D1 expression levels.

Proteomic modulation in breast tumors after metformin exposure: results from a "window of opportunity" trial.

PURPOSE: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). METHODS: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabetic patients with stage 0-III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 %. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. RESULTS: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. CONTROLS: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. CONCLUSIONS: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. CLINICALTRIALS. GOV IDENTIFIER: NCT00930579.