Ligand-mediated targeting represents the cutting edge in precision-guided therapy for several dis... more Ligand-mediated targeting represents the cutting edge in precision-guided therapy for several diseases. Surface engineering of nanomedicines with ligands exhibiting selective or tailored affinity for overexpressed biomolecules of a specific disease may increase therapeutic efficiency and reduce side effects and recurrence. This review focuses on newly developed approaches and strategies to improve treatment and overcome the mechanisms associated with breast cancer resistance.
Thirty-eight strains of Shiga toxin-producing Escherichia coli (STEC) were characterised in terms... more Thirty-eight strains of Shiga toxin-producing Escherichia coli (STEC) were characterised in terms of biochemical properties, enterohaemolysin production and plasmid carriage. A wide variation in the biochemical properties was observed among the STEC, with 14 distinct biotypes identified. Biotype 1 was the most common, found in 29% of the strains. Enterohaemolysin production was detected in 29% of the strains. Most of the bacterial strains (95%) carried one or more plasmids and considerable heterogeneity in size and combinations was observed. Seven distinct plasmid profiles were identified. The most common profile, characterised by the presence of a single plasmid of ~90 kb, was found in 50% of these strains. These data indicate extensive diversity among STEC strains. No correlation was found among biotype, serotype, enterohaemolysin production and plasmid profile.
Supplemental Material, DS1_VET_10.1177_0300985818763711 for DNA Methylation Status of the Estroge... more Supplemental Material, DS1_VET_10.1177_0300985818763711 for DNA Methylation Status of the Estrogen Receptor α Gene in Canine Mammary Tumors by Yara de Oliveira Brandão, Mariana Busato Toledo, Andressa Chequin, Thierry Grima Cristo, Renato Silva Sousa, Edneia Amancio Souza Ramos, and Giseli Klassen in Veterinary Pathology
The enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is a critical engine t... more The enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is a critical engine that supports glucose catabolism. PFKFB3 produces the signaling molecule fructose-2,6-biphosphate (F2,6BP), which activates the second gatekeeper in glycolysis, 6-phosphofructo-1-kinase (PFK-1), and favors the Warburg phenotype. Transcriptional and post-transcriptional processes regulate the abundance and phosphorylation of PFKFB3 in cells, and its activation has been implicated in the progression of several types of cancer. PFKFB3 is important for sustaining glycolysis in the tumorigenesis environment even under unfavorable conditions, thereby promoting metabolic reprogramming, cell proliferation, DNA repair, and drug resistance. Despite its heterogeneous phenotype, breast cancer has unique characteristics that drive the constitutive and inducible expression of PFKFB3 in this opportunistic glycolytic shift. This enzyme is a point of convergence of multiple exogenous and endogenous growth-promoting and oncogenic signaling pathways, especially kinase cascades. The present review summarizes advances in in vitro and in vivo therapy studies that focus on PFKFB3 and the interplay between hormone receptor status and the underlying essential signal transduction system in breast cancer metabolic remodeling.
Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of dea... more Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of death. Despite the discovery of many antineoplastic drugs for BC, the current therapy is not totally efficient. In this study, we investigated the potential of repurposing the well-known diabetes type II drug liraglutide to modulate epigenetic modifications in BC cells lines in vitro and in vivo via Ehrlich mice tumors models. The in vitro results revealed a significant reduction on cell viability, migration, DNMT activity and displayed lower levels of global DNA methylation in BC cell lines after liraglutide treatment. The interaction between liraglutide and the DNMT enzymes resulted in a decrease profile of DNA methylation for the CDH1, ESR1 and ADAM33 gene promoter regions and, consequently, increased their gene and protein expression levels. To elucidate the possible interaction between liraglutide and the DNMT1 protein, we performed an in silico study that indicates liraglutide binding in the catalytic cleft via hydrogen bonds and salt bridges with the interdomain contacts and disturbs the overall enzyme conformation. The in vivo study was also able to reveal that liraglutide and the combined treatment of liraglutide and paclitaxel or methotrexate were effective in reducing tumor growth. Moreover, the modulation of CDH1 and ADAM33 mouse gene expression by DNA demethylation suggests a role for liraglutide in DNMT activity in vivo. Altogether, these results indicate that liraglutide may be further analysed as a new adjuvant treatment for BC.
Jornal Brasileiro de Patologia e Medicina Laboratorial, 2019
Introduction: Breast cancer is the second leading cause of cancer death among women worldwide, an... more Introduction: Breast cancer is the second leading cause of cancer death among women worldwide, and epidemiological studies may help understanding its mechanisms. Objective: To carry out a survey of the number of breast cancer cases diagnosed in a period of six years. Methods: The profile of breast cancers diagnosed in a tertiary hospital in Curitiba was compared with the literature, using a retrospective analysis of ductal/special types and lobular breast carcinoma reports issued between 2008 and 2013. Results: Three hundred twenty-seven (91.6%) cases of ductal/special types carcinoma and 30 (8.4%) cases of lobular carcinoma were diagnosed, totaling 357 samples. From these cases, 27 (7.5%) were carcinoma in situ (20 ductal and seven lobular) and 330 (92.4%) were invasive carcinoma (307 invasive ductal/special types and 23 lobular). The prevalence of breast cancer among women was 99.1% and the majority of patients were older than 50 years of age (67.2%). Regarding the União Internacional de Controle do Câncer/American Joint Committee on Cancer (UICC/AJCC) staging, 49.2% of the ductal/special types tumors were diagnosed in Stages I or II, while 56.6% of lobular carcinomas were diagnosed in Stages II or III/IV. Regarding the Nottingham score, most cases were intermediate grade (43.9%). A total of 61% of the tumors were estrogen receptor positive (ER+) and 54% were progesterone receptor positive (PR+). Moreover, 36.1% presented positive human epidermal growth factor receptor 2 (HER2+), a rate higher than that indicated by the literature. Conclusion: The breast carcinomas evaluated in this study presented a profile similar to that reported in the literature, with some peculiarities inherent to the local pathology service. Nevertheless, the low frequency of in situ cases indicates failure in early diagnosis.
buffered 10% formalin ethanol xylol paraffin hematoxylin and eosin (HE) phosphate buffer (pH 6.5)... more buffered 10% formalin ethanol xylol paraffin hematoxylin and eosin (HE) phosphate buffer (pH 6.5) Griess solution (0.1% N-1-naphthyl-tilediamine, 1% sulfanilamide in 5% H3PO4) saline Triton X-100 0.1%, TMB 18.4 mM dimethylformamide 8% sodium acetate (NaOAc) p-nitrophenyl-N-acetyl--d-glucosamine N-acetyl--d-glucosamine p-nitrofen ketamine hydrochloride xylazine hydrochloride; metothrexate phosphate buffered saline (PBS, 16.5 mM phosphate, 137 mM NaCl, and 2.7 mM KCl) at pH 7.4 a b s t r a c t Polysaccharides are substances that modify the biological response to several stressors. The present study investigated the antitumor activity of the soluble fraction of polysaccharides (SFP), extracted from cabernet franc red wine, in Walker-256 tumor-bearing rats. The monosaccharide composition had a complex mixture, suggesting the presence of arabinoglactans, mannans, and pectins. Treatment with SFP (30 and 60 mg/kg, oral) for 14 days significantly reduced the tumor weight and volume compared with controls. Treatment with 60 mg/kg SFP reduced blood monocytes and neutrophils, reduced the tumor activity of N-acetylglucosaminidase, myeloperoxidase, and nitric oxide, increased blood lymphocytes, and increased the levels of tumor necrosis factor ␣ (TNF-␣) in tumor tissue. Treatment with SFP also induced the expression of the cell necroptosis-related genes Rip1 and Rip3. The antineoplastic effect of SFP appears to be attributable to its action on the immune system by controlling the tumor microenvironment and stimulating TNF-␣ production, which may trigger the necroptosis pathway.
Quiescin Q6/sulfhydryl oxidases (QSOX) are revisited thiol oxidases considered to be involved in ... more Quiescin Q6/sulfhydryl oxidases (QSOX) are revisited thiol oxidases considered to be involved in the oxidative protein folding, cell cycle control and extracellular matrix remodeling. They contain thioredoxin domains and introduce disulfide bonds into proteins and peptides, with the concomitant hydrogen peroxide formation, likely altering the redox environment. Since it is known that several developmental processes are regulated by the redox state, here we assessed if QSOX could have a role during mouse fetal development. For this purpose, an antirecombinant mouse QSOX antibody was produced and characterized. In E 13.5 , E 16.5 fetal tissues, QSOX immunostaining was confined to mesoderm-and ectodermderived tissues, while in P1 neonatal tissues it was slightly extended to some endoderm-derived tissues. QSOX expression, particularly by epithelial tissues, seemed to be developmentally-regulated, increasing with tissue maturation. QSOX was observed in loose connective tissues in all stages analyzed, intra and possibly extracellularly, in agreement with its putative role in oxidative folding and extracellular matrix remodeling. In conclusion, QSOX is expressed in several tissues during mouse development, but preferentially in those derived from mesoderm and ectoderm, suggesting it could be of relevance during developmental processes. Keywords Redox state Á Mouse development Á Quiescin sulfhydryl oxidase Á Mesoderm Kelly F. Portes, Cecília M. Ikegami have contributed equally to this work.
ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains.... more ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains. ADAM family members have different activities, such as proteolysis and adhesion, making them good candidates to mediate the extracellular matrix remodelling ...
Natural products have been recognized as important bioactive compounds on the basis of their wide... more Natural products have been recognized as important bioactive compounds on the basis of their wide biological properties. Here we investigated the antitumor effect and molecular mechanisms of the diterpene Fruticuline A (fruti) from Salvia lachnostachys, in human cancer cell lineages and Solid Ehrlich Carcinoma in mice. Fruti reduced MCF-7 and HepG2 proliferation by the reduction of Cyclin D1 levels and decreased NF-κB gene levels in both cell types. Furthermore, fruti also induced apoptosis in HepG2 cells, reduced Bcl-2 gene expression and induced necroptosis by increasing Ripk in MCF-7 cells. In mice, fruti prevented tumor development and reduced Cyclin D1, Bcl-2 and Rela gene levels, and reduced the p-NF-κB/NF-κB ratio in tumor tissue. Furthermore, fruti induced necrosis and apoptosis, increased N-acetyl-β-D-glucosaminidase and TNF-α levels and reduced IL-10 and Vegf levels in tumor tissue. Collectively, fruti exerts antitumor effects through the inhibition of the NF-κB pathway, r...
The present study investigated the antineoplastic effects of pectic polysaccharides that were ext... more The present study investigated the antineoplastic effects of pectic polysaccharides that were extracted from green sweet pepper (Capsicum annuum [CAP]) in the Ehrlich carcinoma in mice and in human mammary tumor lineages. After the subcutaneous inoculation of 2 × 10 Ehrlich tumor cells, Female Swiss mice received 50, 100, or 150 mg/kg CAP or vehicle orally once daily or methotrexate (2.5 mg/kg, i.p., every 5 days) for 21 days. CAP dose-dependently reduced Ehrlich tumor growth. It also reduced the viability of MCF-7, MDA-MB-231, and MDA-MB-436 human mammary cell lineages. Treatment with CAP reduced the gene expression of vascular endothelial growth factor in vivo and in vitro, reduced vessel areas of the tumors, and induced necrosis in Ehrlich solid tumors. CAP treatment significantly increased Interleukin-6 in tumors. The antineoplastic effect of CAP appears to depend on the regulation of inflammation and angiogenesis. Further studies are encouraged to better understand the CAP pote...
Estrogen receptor α (ERα) has an important role in mammary carcinogenesis, prognosis, and treatme... more Estrogen receptor α (ERα) has an important role in mammary carcinogenesis, prognosis, and treatment. In human and canine mammary cancer, the most aggressive tumors show loss of ERα expression, which in human breast cancer has been attributed to methylation of the cytosine followed by guanine (CpG) island within the estrogen receptor α gene ( ESR1) promoter. This study aimed to investigate the role of ESR1 CpG island (CGI) methylation in ERα expression in canine mammary tumors. Twenty-one canine mammary samples were sorted into three groups: malignant tumor (n = 9), benign tumor (n = 8), and normal gland (n = 4). Immunohistochemical analysis and reverse-transcription quantitative real-time PCR were performed to assess ERα expression and ESR1 mRNA levels. The methylation status was determined using sodium-bisulfite-treated DNA sequencing. All normal mammary glands and benign tumors showed high ERα expression (score range, 5-8). Six of the nine malignant tumors did not show ERα express...
Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death among... more Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death among women worldwide. Metastasis remains a major challenge for the clinical management and prognosis of patients with cancer. The metalloprotease MMP-9 plays a critical role in the first step of metastasis through extracellular matrix degradation. In this study, our goal was to determine the effect of epigenetic mechanisms in the promoter and intragenic region of this gene and to correlate it to the levels of expression of MMP9 in breast cancer cell lines. We have identified that MMP9 was highly expressed in the breast cancer cell lines MCF7 and MDA-MB-436 after 5-aza-2'-deoxycytidine (5-azadC) treatment. Sequencing of the promoter region as well as the CGI intronic CpG islands showed a specific sequence in CGI2, between CpGs 12-30 that was demethylated after 5-azadC treatment. This specific region was studied in breast cancer samples that revealed similar results with demethylation in p...
The present study evaluated the in vivo antitumor effects and toxicity of a new Ru(II) compound, ... more The present study evaluated the in vivo antitumor effects and toxicity of a new Ru(II) compound, cis-(Ru[phen]2[ImH]2)(2+) (also called RuphenImH [RuC]), against Walker-256 carcinosarcoma in rats. After subcutaneous inoculation of Walker-256 cells in the right pelvic limb, male Wistar rats received 5 or 10mgkg(-1) RuC orally or intraperitoneally (i.p.) every 3 days for 13 days. A positive control group (2mgkg(-1) cisplatin) and negative control group (vehicle) were also used. Tumor progression was checked daily. After treatment, tumor weight, plasma biochemistry, hematology, oxidative stress, histology, and tumor cell respiration were evaluated. RuC was effective against tumors when administered i.p. but not orally. The highest i.p. dose of RuC (10mgkg(-1)) significantly reduced tumor volume and weight, induced oxidative stress in tumor tissue, reduced the respiration of tumor cells, and induced necrosis but did not induce apoptosis in the tumor. No clinical signs of toxicity or dea...
Revista da Associacao Medica Brasileira (1992), 2016
Breast cancer is the most cause of death, and approximately 90% of these deaths are due to metast... more Breast cancer is the most cause of death, and approximately 90% of these deaths are due to metastases. Matrix metalloproteinase-2 (MMP-2) gelatinase activity is able to degrade a major constituent of the tumor microenvironment, type IV collagen. Two well-established proteins used as markers in clinical practice for breast cancer are the receptors for estrogen (ER) and progesterone (PR). Although the presence of these receptors has been associated with a better prognosis, loss of these proteins can occur during tumor progression, with subsequent resistance to hormone therapy. To study the correlation among MMP-2, ER, and PR, as well as the establishment of the metastatic process in primary breast tumors. Breast cancer samples (n=44) were analyzed by immunohistochemistry for MMP-2, ER, and PR. We observed that 90% of patients who had metastases and died showed positive staining for MMP-2 (p=0.0082 for both). Using Kaplan-Meier analysis, we found that negative ER patients who were also...
Journal of clinical laboratory analysis, Jan 24, 2016
The glucokinase regulatory protein (GCKR) regulates the activity of the glucokinase (GCK), which ... more The glucokinase regulatory protein (GCKR) regulates the activity of the glucokinase (GCK), which plays a key role in glucose homeostasis. Genetic variants in GCK have been associated with diabetes and gestational diabetes (GDM). Due to the relationship between GCKRP and GCK, polymorphisms in GCKR are also candidates for genetic association with GDM. The aim of this study was to evaluate the association between the GCKR rs780094 polymorphism and GDM in a Brazilian population. 252 unrelated Euro-Brazilian pregnant women were classified as control (healthy pregnant women, n = 125) and GDM (pregnant women with GDM, n = 127) age-matched groups. Clinical and anthropometric data were obtained from all subjects. The GCKR rs780094 polymorphism was genotyped using fluorescent probes (TaqMan(®) , code C_2862873_10). Both groups were in Hardy-Weinberg equilibrium. The GCKR rs780094 polymorphism was associated with GDM in codominant and dominant models (P = 0.022 and P = 0.010, respectively). Th...
Ligand-mediated targeting represents the cutting edge in precision-guided therapy for several dis... more Ligand-mediated targeting represents the cutting edge in precision-guided therapy for several diseases. Surface engineering of nanomedicines with ligands exhibiting selective or tailored affinity for overexpressed biomolecules of a specific disease may increase therapeutic efficiency and reduce side effects and recurrence. This review focuses on newly developed approaches and strategies to improve treatment and overcome the mechanisms associated with breast cancer resistance.
Thirty-eight strains of Shiga toxin-producing Escherichia coli (STEC) were characterised in terms... more Thirty-eight strains of Shiga toxin-producing Escherichia coli (STEC) were characterised in terms of biochemical properties, enterohaemolysin production and plasmid carriage. A wide variation in the biochemical properties was observed among the STEC, with 14 distinct biotypes identified. Biotype 1 was the most common, found in 29% of the strains. Enterohaemolysin production was detected in 29% of the strains. Most of the bacterial strains (95%) carried one or more plasmids and considerable heterogeneity in size and combinations was observed. Seven distinct plasmid profiles were identified. The most common profile, characterised by the presence of a single plasmid of ~90 kb, was found in 50% of these strains. These data indicate extensive diversity among STEC strains. No correlation was found among biotype, serotype, enterohaemolysin production and plasmid profile.
Supplemental Material, DS1_VET_10.1177_0300985818763711 for DNA Methylation Status of the Estroge... more Supplemental Material, DS1_VET_10.1177_0300985818763711 for DNA Methylation Status of the Estrogen Receptor α Gene in Canine Mammary Tumors by Yara de Oliveira Brandão, Mariana Busato Toledo, Andressa Chequin, Thierry Grima Cristo, Renato Silva Sousa, Edneia Amancio Souza Ramos, and Giseli Klassen in Veterinary Pathology
The enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is a critical engine t... more The enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is a critical engine that supports glucose catabolism. PFKFB3 produces the signaling molecule fructose-2,6-biphosphate (F2,6BP), which activates the second gatekeeper in glycolysis, 6-phosphofructo-1-kinase (PFK-1), and favors the Warburg phenotype. Transcriptional and post-transcriptional processes regulate the abundance and phosphorylation of PFKFB3 in cells, and its activation has been implicated in the progression of several types of cancer. PFKFB3 is important for sustaining glycolysis in the tumorigenesis environment even under unfavorable conditions, thereby promoting metabolic reprogramming, cell proliferation, DNA repair, and drug resistance. Despite its heterogeneous phenotype, breast cancer has unique characteristics that drive the constitutive and inducible expression of PFKFB3 in this opportunistic glycolytic shift. This enzyme is a point of convergence of multiple exogenous and endogenous growth-promoting and oncogenic signaling pathways, especially kinase cascades. The present review summarizes advances in in vitro and in vivo therapy studies that focus on PFKFB3 and the interplay between hormone receptor status and the underlying essential signal transduction system in breast cancer metabolic remodeling.
Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of dea... more Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of death. Despite the discovery of many antineoplastic drugs for BC, the current therapy is not totally efficient. In this study, we investigated the potential of repurposing the well-known diabetes type II drug liraglutide to modulate epigenetic modifications in BC cells lines in vitro and in vivo via Ehrlich mice tumors models. The in vitro results revealed a significant reduction on cell viability, migration, DNMT activity and displayed lower levels of global DNA methylation in BC cell lines after liraglutide treatment. The interaction between liraglutide and the DNMT enzymes resulted in a decrease profile of DNA methylation for the CDH1, ESR1 and ADAM33 gene promoter regions and, consequently, increased their gene and protein expression levels. To elucidate the possible interaction between liraglutide and the DNMT1 protein, we performed an in silico study that indicates liraglutide binding in the catalytic cleft via hydrogen bonds and salt bridges with the interdomain contacts and disturbs the overall enzyme conformation. The in vivo study was also able to reveal that liraglutide and the combined treatment of liraglutide and paclitaxel or methotrexate were effective in reducing tumor growth. Moreover, the modulation of CDH1 and ADAM33 mouse gene expression by DNA demethylation suggests a role for liraglutide in DNMT activity in vivo. Altogether, these results indicate that liraglutide may be further analysed as a new adjuvant treatment for BC.
Jornal Brasileiro de Patologia e Medicina Laboratorial, 2019
Introduction: Breast cancer is the second leading cause of cancer death among women worldwide, an... more Introduction: Breast cancer is the second leading cause of cancer death among women worldwide, and epidemiological studies may help understanding its mechanisms. Objective: To carry out a survey of the number of breast cancer cases diagnosed in a period of six years. Methods: The profile of breast cancers diagnosed in a tertiary hospital in Curitiba was compared with the literature, using a retrospective analysis of ductal/special types and lobular breast carcinoma reports issued between 2008 and 2013. Results: Three hundred twenty-seven (91.6%) cases of ductal/special types carcinoma and 30 (8.4%) cases of lobular carcinoma were diagnosed, totaling 357 samples. From these cases, 27 (7.5%) were carcinoma in situ (20 ductal and seven lobular) and 330 (92.4%) were invasive carcinoma (307 invasive ductal/special types and 23 lobular). The prevalence of breast cancer among women was 99.1% and the majority of patients were older than 50 years of age (67.2%). Regarding the União Internacional de Controle do Câncer/American Joint Committee on Cancer (UICC/AJCC) staging, 49.2% of the ductal/special types tumors were diagnosed in Stages I or II, while 56.6% of lobular carcinomas were diagnosed in Stages II or III/IV. Regarding the Nottingham score, most cases were intermediate grade (43.9%). A total of 61% of the tumors were estrogen receptor positive (ER+) and 54% were progesterone receptor positive (PR+). Moreover, 36.1% presented positive human epidermal growth factor receptor 2 (HER2+), a rate higher than that indicated by the literature. Conclusion: The breast carcinomas evaluated in this study presented a profile similar to that reported in the literature, with some peculiarities inherent to the local pathology service. Nevertheless, the low frequency of in situ cases indicates failure in early diagnosis.
buffered 10% formalin ethanol xylol paraffin hematoxylin and eosin (HE) phosphate buffer (pH 6.5)... more buffered 10% formalin ethanol xylol paraffin hematoxylin and eosin (HE) phosphate buffer (pH 6.5) Griess solution (0.1% N-1-naphthyl-tilediamine, 1% sulfanilamide in 5% H3PO4) saline Triton X-100 0.1%, TMB 18.4 mM dimethylformamide 8% sodium acetate (NaOAc) p-nitrophenyl-N-acetyl--d-glucosamine N-acetyl--d-glucosamine p-nitrofen ketamine hydrochloride xylazine hydrochloride; metothrexate phosphate buffered saline (PBS, 16.5 mM phosphate, 137 mM NaCl, and 2.7 mM KCl) at pH 7.4 a b s t r a c t Polysaccharides are substances that modify the biological response to several stressors. The present study investigated the antitumor activity of the soluble fraction of polysaccharides (SFP), extracted from cabernet franc red wine, in Walker-256 tumor-bearing rats. The monosaccharide composition had a complex mixture, suggesting the presence of arabinoglactans, mannans, and pectins. Treatment with SFP (30 and 60 mg/kg, oral) for 14 days significantly reduced the tumor weight and volume compared with controls. Treatment with 60 mg/kg SFP reduced blood monocytes and neutrophils, reduced the tumor activity of N-acetylglucosaminidase, myeloperoxidase, and nitric oxide, increased blood lymphocytes, and increased the levels of tumor necrosis factor ␣ (TNF-␣) in tumor tissue. Treatment with SFP also induced the expression of the cell necroptosis-related genes Rip1 and Rip3. The antineoplastic effect of SFP appears to be attributable to its action on the immune system by controlling the tumor microenvironment and stimulating TNF-␣ production, which may trigger the necroptosis pathway.
Quiescin Q6/sulfhydryl oxidases (QSOX) are revisited thiol oxidases considered to be involved in ... more Quiescin Q6/sulfhydryl oxidases (QSOX) are revisited thiol oxidases considered to be involved in the oxidative protein folding, cell cycle control and extracellular matrix remodeling. They contain thioredoxin domains and introduce disulfide bonds into proteins and peptides, with the concomitant hydrogen peroxide formation, likely altering the redox environment. Since it is known that several developmental processes are regulated by the redox state, here we assessed if QSOX could have a role during mouse fetal development. For this purpose, an antirecombinant mouse QSOX antibody was produced and characterized. In E 13.5 , E 16.5 fetal tissues, QSOX immunostaining was confined to mesoderm-and ectodermderived tissues, while in P1 neonatal tissues it was slightly extended to some endoderm-derived tissues. QSOX expression, particularly by epithelial tissues, seemed to be developmentally-regulated, increasing with tissue maturation. QSOX was observed in loose connective tissues in all stages analyzed, intra and possibly extracellularly, in agreement with its putative role in oxidative folding and extracellular matrix remodeling. In conclusion, QSOX is expressed in several tissues during mouse development, but preferentially in those derived from mesoderm and ectoderm, suggesting it could be of relevance during developmental processes. Keywords Redox state Á Mouse development Á Quiescin sulfhydryl oxidase Á Mesoderm Kelly F. Portes, Cecília M. Ikegami have contributed equally to this work.
ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains.... more ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains. ADAM family members have different activities, such as proteolysis and adhesion, making them good candidates to mediate the extracellular matrix remodelling ...
Natural products have been recognized as important bioactive compounds on the basis of their wide... more Natural products have been recognized as important bioactive compounds on the basis of their wide biological properties. Here we investigated the antitumor effect and molecular mechanisms of the diterpene Fruticuline A (fruti) from Salvia lachnostachys, in human cancer cell lineages and Solid Ehrlich Carcinoma in mice. Fruti reduced MCF-7 and HepG2 proliferation by the reduction of Cyclin D1 levels and decreased NF-κB gene levels in both cell types. Furthermore, fruti also induced apoptosis in HepG2 cells, reduced Bcl-2 gene expression and induced necroptosis by increasing Ripk in MCF-7 cells. In mice, fruti prevented tumor development and reduced Cyclin D1, Bcl-2 and Rela gene levels, and reduced the p-NF-κB/NF-κB ratio in tumor tissue. Furthermore, fruti induced necrosis and apoptosis, increased N-acetyl-β-D-glucosaminidase and TNF-α levels and reduced IL-10 and Vegf levels in tumor tissue. Collectively, fruti exerts antitumor effects through the inhibition of the NF-κB pathway, r...
The present study investigated the antineoplastic effects of pectic polysaccharides that were ext... more The present study investigated the antineoplastic effects of pectic polysaccharides that were extracted from green sweet pepper (Capsicum annuum [CAP]) in the Ehrlich carcinoma in mice and in human mammary tumor lineages. After the subcutaneous inoculation of 2 × 10 Ehrlich tumor cells, Female Swiss mice received 50, 100, or 150 mg/kg CAP or vehicle orally once daily or methotrexate (2.5 mg/kg, i.p., every 5 days) for 21 days. CAP dose-dependently reduced Ehrlich tumor growth. It also reduced the viability of MCF-7, MDA-MB-231, and MDA-MB-436 human mammary cell lineages. Treatment with CAP reduced the gene expression of vascular endothelial growth factor in vivo and in vitro, reduced vessel areas of the tumors, and induced necrosis in Ehrlich solid tumors. CAP treatment significantly increased Interleukin-6 in tumors. The antineoplastic effect of CAP appears to depend on the regulation of inflammation and angiogenesis. Further studies are encouraged to better understand the CAP pote...
Estrogen receptor α (ERα) has an important role in mammary carcinogenesis, prognosis, and treatme... more Estrogen receptor α (ERα) has an important role in mammary carcinogenesis, prognosis, and treatment. In human and canine mammary cancer, the most aggressive tumors show loss of ERα expression, which in human breast cancer has been attributed to methylation of the cytosine followed by guanine (CpG) island within the estrogen receptor α gene ( ESR1) promoter. This study aimed to investigate the role of ESR1 CpG island (CGI) methylation in ERα expression in canine mammary tumors. Twenty-one canine mammary samples were sorted into three groups: malignant tumor (n = 9), benign tumor (n = 8), and normal gland (n = 4). Immunohistochemical analysis and reverse-transcription quantitative real-time PCR were performed to assess ERα expression and ESR1 mRNA levels. The methylation status was determined using sodium-bisulfite-treated DNA sequencing. All normal mammary glands and benign tumors showed high ERα expression (score range, 5-8). Six of the nine malignant tumors did not show ERα express...
Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death among... more Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death among women worldwide. Metastasis remains a major challenge for the clinical management and prognosis of patients with cancer. The metalloprotease MMP-9 plays a critical role in the first step of metastasis through extracellular matrix degradation. In this study, our goal was to determine the effect of epigenetic mechanisms in the promoter and intragenic region of this gene and to correlate it to the levels of expression of MMP9 in breast cancer cell lines. We have identified that MMP9 was highly expressed in the breast cancer cell lines MCF7 and MDA-MB-436 after 5-aza-2'-deoxycytidine (5-azadC) treatment. Sequencing of the promoter region as well as the CGI intronic CpG islands showed a specific sequence in CGI2, between CpGs 12-30 that was demethylated after 5-azadC treatment. This specific region was studied in breast cancer samples that revealed similar results with demethylation in p...
The present study evaluated the in vivo antitumor effects and toxicity of a new Ru(II) compound, ... more The present study evaluated the in vivo antitumor effects and toxicity of a new Ru(II) compound, cis-(Ru[phen]2[ImH]2)(2+) (also called RuphenImH [RuC]), against Walker-256 carcinosarcoma in rats. After subcutaneous inoculation of Walker-256 cells in the right pelvic limb, male Wistar rats received 5 or 10mgkg(-1) RuC orally or intraperitoneally (i.p.) every 3 days for 13 days. A positive control group (2mgkg(-1) cisplatin) and negative control group (vehicle) were also used. Tumor progression was checked daily. After treatment, tumor weight, plasma biochemistry, hematology, oxidative stress, histology, and tumor cell respiration were evaluated. RuC was effective against tumors when administered i.p. but not orally. The highest i.p. dose of RuC (10mgkg(-1)) significantly reduced tumor volume and weight, induced oxidative stress in tumor tissue, reduced the respiration of tumor cells, and induced necrosis but did not induce apoptosis in the tumor. No clinical signs of toxicity or dea...
Revista da Associacao Medica Brasileira (1992), 2016
Breast cancer is the most cause of death, and approximately 90% of these deaths are due to metast... more Breast cancer is the most cause of death, and approximately 90% of these deaths are due to metastases. Matrix metalloproteinase-2 (MMP-2) gelatinase activity is able to degrade a major constituent of the tumor microenvironment, type IV collagen. Two well-established proteins used as markers in clinical practice for breast cancer are the receptors for estrogen (ER) and progesterone (PR). Although the presence of these receptors has been associated with a better prognosis, loss of these proteins can occur during tumor progression, with subsequent resistance to hormone therapy. To study the correlation among MMP-2, ER, and PR, as well as the establishment of the metastatic process in primary breast tumors. Breast cancer samples (n=44) were analyzed by immunohistochemistry for MMP-2, ER, and PR. We observed that 90% of patients who had metastases and died showed positive staining for MMP-2 (p=0.0082 for both). Using Kaplan-Meier analysis, we found that negative ER patients who were also...
Journal of clinical laboratory analysis, Jan 24, 2016
The glucokinase regulatory protein (GCKR) regulates the activity of the glucokinase (GCK), which ... more The glucokinase regulatory protein (GCKR) regulates the activity of the glucokinase (GCK), which plays a key role in glucose homeostasis. Genetic variants in GCK have been associated with diabetes and gestational diabetes (GDM). Due to the relationship between GCKRP and GCK, polymorphisms in GCKR are also candidates for genetic association with GDM. The aim of this study was to evaluate the association between the GCKR rs780094 polymorphism and GDM in a Brazilian population. 252 unrelated Euro-Brazilian pregnant women were classified as control (healthy pregnant women, n = 125) and GDM (pregnant women with GDM, n = 127) age-matched groups. Clinical and anthropometric data were obtained from all subjects. The GCKR rs780094 polymorphism was genotyped using fluorescent probes (TaqMan(®) , code C_2862873_10). Both groups were in Hardy-Weinberg equilibrium. The GCKR rs780094 polymorphism was associated with GDM in codominant and dominant models (P = 0.022 and P = 0.010, respectively). Th...
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