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1.
Biomed Pharmacother ; 165: 115179, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37481927

ABSTRACT

Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers worldwide, primarily due to its robust desmoplastic stroma and immunosuppressive tumor microenvironment (TME), which facilitate tumor progression and metastasis. In addition, fibrous tissue leads to sparse vasculature, high interstitial fluid pressure, and hypoxia, thereby hindering effective systemic drug delivery and immune cell infiltration. Thus, remodeling the TME to enhance tumor perfusion, increase drug retention, and reverse immunosuppression has become a key therapeutic strategy. In recent years, targeting epigenetic pathways has emerged as a promising approach to overcome tumor immunosuppression and cancer progression. Moreover, the progress in nanotechnology has provided new opportunities for enhancing the efficacy of conventional and epigenetic drugs. Nano-based drug delivery systems (NDDSs) offer several advantages, including improved drug pharmacokinetics, enhanced tumor penetration, and reduced systemic toxicity. Smart NDDSs enable precise targeting of stromal components and augment the effectiveness of immunotherapy through multiple drug delivery options. This review offers an overview of the latest nano-based approaches developed to achieve superior therapeutic efficacy and overcome drug resistance. We specifically focus on the TME and epigenetic-targeted therapies in the context of PDAC, discussing the advantages and limitations of current strategies while highlighting promising new developments. By emphasizing the immense potential of NDDSs in improving therapeutic outcomes in PDAC, our review paves the way for future research in this rapidly evolving field.


Subject(s)
Carcinoma, Pancreatic Ductal , Pancreatic Neoplasms , Humans , Nanomedicine , Nanoparticle Drug Delivery System , Pancreatic Neoplasms/drug therapy , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/pathology , Carcinoma, Pancreatic Ductal/drug therapy , Carcinoma, Pancreatic Ductal/genetics , Carcinoma, Pancreatic Ductal/pathology , Tumor Microenvironment/genetics , Pancreatic Neoplasms
2.
Cancers (Basel) ; 15(10)2023 May 19.
Article in English | MEDLINE | ID: mdl-37345168

ABSTRACT

Gastrointestinal (GI) cancers are malignancies that develop within the digestive system and account for one in four cancer cases according to WHO data [...].

3.
Int J Mol Sci ; 23(24)2022 Dec 07.
Article in English | MEDLINE | ID: mdl-36555088

ABSTRACT

Genetic aberrations, including chromosomal rearrangements, loss or amplification of DNA, and point mutations, are major elements of cancer development [...].


Subject(s)
Epigenesis, Genetic , Neoplasms , Humans , Chromosome Aberrations , Neoplasms/genetics , Neoplasms/therapy , Point Mutation , DNA
4.
Crit Rev Oncol Hematol ; 180: 103865, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36334880

ABSTRACT

Pancreatic neuroendocrine neoplasms (pNENs) are rare cancers with broad challenges for their management. The main clinical obstacles are the high rate of patients diagnosed at advanced stages, lack of prognostic markers for early detection of disease recurrence in resected patients, significant limitations in identifying those who will benefit from adjuvant therapy, and timely recognition of treatment response. Therefore, the discovery of new prognostic and predictive markers is necessary for patient stratification and clinical management. Liquid biopsy, which has revolutionized the field of clinical oncology, is extremely under-investigated in pNENs. This review highlights its potential and the recent advances in related technologies, as candidates for the delivery of the new tools that can help to refine pNEN diagnosis and to personalize treatment. In addition, the opportunities and limitations of available preclinical research models with regard to biomarker research are discussed in light of pNEN clinical needs.


Subject(s)
Neuroendocrine Tumors , Pancreatic Neoplasms , Humans , Neuroendocrine Tumors/diagnosis , Neuroendocrine Tumors/therapy , Neuroendocrine Tumors/pathology , Pancreatic Neoplasms/diagnosis , Pancreatic Neoplasms/therapy , Pancreatic Neoplasms/pathology , Neoplasm Recurrence, Local , Liquid Biopsy , Prognosis
5.
Oncol Lett ; 22(6): 820, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34691247

ABSTRACT

Previous studies have reported that CD44 variant 6 (CD44v6) and metastasis-associated protein 1 (MTA1) are contributing factors to cancer progression. The present study aimed to evaluate the expression profiles for associations with patients' demographic data, clinicopathological characteristics, the presence of partial epithelial-to-mesenchymal transition (pEMT), metastatic potential based on the presence of CK20+ CEA+ CXCR4+ circulating tumor cells (CTCs) and prognosis (median follow-up, 45 months). Thus, frozen tissue samples from 31 patients with stage I-III colorectal cancer (CRC), 15 benign colorectal polyps and seven normal colorectal tissues were analyzed to detect membranous (m)CD44v6 and MTA1 expression via flow cytometry. The results demonstrated that the mCD44v6 and MTA1 expression profiles were significantly correlated (rs=+0.786, P<0.001). Notably, MTA1 expression was not associated with any of the clinicopathological characteristics assessed. The percentage of mCD44v6-positive cells within tumors was higher in the right-sided cancer lesions (P=0.014), suggesting that proximal and distal CRCs are distinct clinicopathological entities. Furthermore, downregulated mCD44v6 expression was significantly associated with the presence of CTCs (P=0.017). This association was stronger for pEMT (co-expression of N- and E-cadherin mRNAs) primary lesions (P=0.009). In addition, patients with CRC with low levels of mCD44v6 had unfavorable survival outcomes (P=0.037). Taken together, these results suggest that targeted analysis of membranous CD44v6 as opposed to membranous-cytoplasmic expression is important in determining the prognosis of patients with CRC. Furthermore, downregulated mCD44v6 expression in malignancies presenting CTCs reinforces the importance of tumor-stroma reciprocal influence during the metastatic process and encourages the assessment of relevant therapeutic strategies.

6.
Cancers (Basel) ; 13(18)2021 Sep 21.
Article in English | MEDLINE | ID: mdl-34572959

ABSTRACT

Cell senescence constitutes a physiological process that serves as protection from malignant transformation of cells. However, recent scientific discoveries also identify cell senescence as pivotal in hepatocellular cancer (HCC) biology. The review herein aimed to accumulate evidence on senescence as a mediator of HCC occurrence in hepatitis B (HBV), C (HCV) virus infections, and non-alcoholic fatty liver disease (NAFLD). In HBV infection, the carcinogenic HBV X protein frequently mutates during chronic infection, and subsequently exhibits different effects on senescence. In HCV infection, senescent non-functional T-cells do not effectively clear pre-malignant hepatocytes. Furthermore, the HCV Core protein inhibits the occurrence of normal stress-induced hepatocyte senescence, allowing damaged cells to maintain their proliferative potential. In NAFLD-mediated HCC, current data point towards the gut microbiome and hepatic stellate cell senescence. Additionally, senescence contributes in the development of resistance in targeted therapies, such as sorafenib. Finally, the promising role of senotherapeutics in HCC was also explored. Overall, although we may still be at a primitive stage in fully unraveling the role of senescence in cancer, it seems that understanding and harnessing senescence may have the potential to revolutionize the way we treat hepatocellular cancer.

7.
In Vivo ; 35(1): 333-339, 2021.
Article in English | MEDLINE | ID: mdl-33402482

ABSTRACT

BACKGROUND/AIM: The mechanisms underlying the contribution of the heparan sulfate proteoglycan syndecan-1 to liver tissue injury and to crucial biological processes, such as fibrogenesis, remain to be elucidated. Therefore, we investigated the immunohistochemical expression of syndecan-1 in chronic liver diseases (CLDs) and its probable role in hepatic fibrosis. MATERIALS AND METHODS: Immunohistochemistry was performed on formalin-fixed, paraffin-embedded tissue sections of biopsy material obtained from 128 patients diagnosed with CLDs. The correlation between syndecan-1 expression and the stage of fibrosis was investigated. RESULTS: According to the severity of fibrosis, cases were categorized into three groups: early fibrosis; intermediate fibrosis; advanced fibrosis. Syndecan-1 expression was significantly enhanced in advanced fibrosis compared to early (p<0.012) and intermediate (p<0.003) fibrosis. CONCLUSION: In CLDs, syndecan-1 immunohisto-chemical overexpression was found to be positively correlated with the severity of fibrosis, suggesting its probable role in hepatic fibrogenesis.


Subject(s)
Membrane Glycoproteins , Syndecan-1 , Humans , Immunohistochemistry , Liver Cirrhosis/genetics , Syndecan-1/genetics
8.
Cancer Biomark ; 25(2): 141-149, 2019.
Article in English | MEDLINE | ID: mdl-31104007

ABSTRACT

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) constitutes the third most frequent head and neck cancer. Several tissue biomarkers have been studied for their prognostic significance in LSCC. OBJECTIVE: To investigate the prognostic significance of BCL2L12, a new member of the BCL2 family, in primary LSCC along with well-examined biomarkers such as BCL2 and BAX. METHODS: Cancerous tissue specimens of patients with primary LSCC were collected during 2005 and 2012 as pretreatment tissue biopsy. The specimens were immunohistochemically evaluated for the protein expression of BCL2L12, BCL2 and BAX. Kaplan-Meier survival curves and Cox proportional hazard regression models were performed to evaluate prognosis. RESULTS: In the study cohort of 78 patients with primary LSCC, Kaplan-Meier survival curves demonstrated that advanced-stage LSCC patients with BCL2L12-positive tumors had significantly higher OS time in comparison with advanced-stage LSCC patients with BCL2L12-negative tumors (p= 0.014). Also, advanced-stage LSCC patients with BCL2L12-positive tumors had significantly lower risk of death from LSCC compared to advanced-stage LSCC patients with BCL2L12-negative tumors (HR = 0.228, 95%CI = 0.063-0.833, p= 0.025). CONCLUSIONS: BCL2L12 protein expression could be used as a favorable prognostic tissue biomarker in patients with primary advanced-stage LSCC. On the contrary, BCL2 and BAX did not correlate with prognosis in patients with primary LSCC.


Subject(s)
Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/mortality , Gene Expression Regulation, Neoplastic , Laryngeal Neoplasms/genetics , Laryngeal Neoplasms/mortality , Muscle Proteins/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor , Carcinoma, Squamous Cell/pathology , Female , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Laryngeal Neoplasms/pathology , Male , Middle Aged , Muscle Proteins/metabolism , Neoplasm Grading , Neoplasm Staging , Prognosis , Proportional Hazards Models , Proto-Oncogene Proteins c-bcl-2/metabolism , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolism
9.
Am J Otolaryngol ; 40(4): 487-493, 2019.
Article in English | MEDLINE | ID: mdl-30979652

ABSTRACT

PURPOSE: Laryngeal squamous cell carcinoma (LSCC), a common type of head and neck cancer, is associated with high rates of metastasis and recurrence. In this study, we investigated the potential combinatorial prognostic value of NOTCH1, Vimentin (VIM), and Metastasis-associated 1 (MTA1) protein in LSCC, using immunohistochemistry. MATERIALS AND METHODS: Tissue specimens from 69 patients with LSCC were immunohistochemically evaluated for the protein expression of NOTCH1, VIM, and MTA1. Then, biostatistical analysis was performed, in order to assess the prognostic value of the expression of each one of these proteins. RESULTS: NOTCH1 expression status was not a significant prognosticator in LSCC, as shown in Kaplan-Meier survival analysis. On the contrary, both VIM and MTA1 seem to have an important prognostic potential, independently of TNM staging and histological grade of the tumor. In fact, positive VIM expression was shown to predict patients' relapse and poor outcome regarding patients' overall survival, in contrast with MTA1, the positive expression of which predicts higher disease-free survival (DFS) and overall survival (OS) rates in LSCC. CONCLUSIONS: VIM and MTA1 constitute potential tumor biomarkers in LSCC and could be integrated into a multiparametric prognostic model. Undoubtedly, their prognostic value needs further validation in larger cohorts of LSCC patients.


Subject(s)
Gene Expression , Laryngeal Neoplasms/genetics , Laryngeal Neoplasms/metabolism , Repressor Proteins/genetics , Repressor Proteins/metabolism , Squamous Cell Carcinoma of Head and Neck/genetics , Squamous Cell Carcinoma of Head and Neck/metabolism , Trans-Activators/genetics , Trans-Activators/metabolism , Vimentin/genetics , Vimentin/metabolism , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Female , Humans , Immunohistochemistry , Laryngeal Neoplasms/diagnosis , Laryngeal Neoplasms/mortality , Male , Middle Aged , Prognosis , Receptor, Notch1/genetics , Receptor, Notch1/metabolism , Squamous Cell Carcinoma of Head and Neck/diagnosis , Squamous Cell Carcinoma of Head and Neck/mortality , Survival Rate
10.
Pathol Res Pract ; 214(12): 2004-2010, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30297114

ABSTRACT

BACKGROUND: Nasal polyposis (NP) and sinonasal inverted papillomas (SIP) are considered benign lesions capable of recurrence or malignant transformation although not with the same prevalence. Since fluctuations of Caveolin-1 and Notch-1 proteins expression have been reported in many pathologies, the current study aimed to investigate their involvement in the epithelial transformation observed in SIPs compared to NP. METHODS: Immunohistochemical expression of Caveolin-1 and Notch-1 proteins was assessed in 104 patients with sinonasal lesions (45 NP, 45 SIP and 14 NP with SIP), semiquantively (percentage times intensity). Proteins expression profiles were evaluated statistically for their correlation with patients demographic and clinicopathological variables (grade of dysplasia, inflammation, recurrence) as well as with markers of proliferation (Ki67) and apoptosis (7-AAD) as determined by flow cytometry analysis. RESULTS: SIP lesions presented increased Caveolin-1 immunopositivity compared to NP (62.2%, vs 40.9%; p = 0.045). Cytoplasmic staining was observed only in epithelium's basal and suprabasal layers. Caveolin-1 positivity was not related to Ki67 expression, apoptosis, inflammation or dysplasia, eventhough 81.8% of highly immunopositive lesions were dysplastic (p = 0.03). Also, smokers presented significantly increased immunopositivy (p = 0.03). In contrast SIP lesions presented reduced Notch-1 expression compared to NP (68.9% vs 100%; p < 0.001). Dysplastic lesions presented low Notch-1 immunopositivity (p < 0.001). Enhancement of Notch-1 gene expression was also associated with inflammation. CONCLUSIONS: The herein presented data suggest that the expression profiles of Caveolin-1 and Notch-1 proteins in sinonasal pathologies are distinctive and that could be explored as potential targets for the development of alternative therapeutic approaches.


Subject(s)
Caveolin 1/metabolism , Nasal Polyps/metabolism , Nose Neoplasms/metabolism , Papilloma, Inverted/metabolism , Receptor, Notch1/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Apoptosis/physiology , Cell Proliferation/physiology , Female , Humans , Male , Middle Aged , Nasal Polyps/pathology , Nose Neoplasms/pathology , Papilloma, Inverted/pathology , Young Adult
11.
Shock ; 50(2): 199-208, 2018 08.
Article in English | MEDLINE | ID: mdl-28957875

ABSTRACT

The present study focuses on the profile of "endogeneous" caveolin-1 protein in septic lung (CLP model).Caveolin-1, CD25, pP38, pAkt, and 14-3-3b protein expression profiles were studied using flow cytometry and immunohistochemistry 6, 12, 24, 36, and 48 h after sepsis induction. Cell viability was determined by 7-AAD staining and fibrosis by Masson trichrome stain. The effect of protein C zymogen concentrate (PC) on caveolin-1 expression was also investigated given that PC, once dissociated from caveolin-1, elicits a PAR-1-mediated protective signaling by forming a complex with endothelial protein C receptor (EPCR).CLP treatment increased lung inflammation and cell apoptosis. Fibrosis was apparent in vessels and alveoli. Caveolin-1+ cells presented reduced protein expression, especially 12 h post-CLP (P = 0.002). Immunohistochemistry revealed caveolin-1 positive expression mainly in regions with strong inflammatory reaction. Early induction of pP38+ cell population (P = 0.014) and gradual increase of CD25+ cells were also observed. Alternations in 14-3-3b expression related to apoptosis were apparent and accompanied by increased AKT phosphorylation activity late during sepsis progression.After PC administration, cell apoptosis was reduced (P = 0.004) and both the percentile and expression intensity of caveolin-1 positive cells were compromised (P = 0.009 and P = 0.027, respectively). 14-3-3b, CD25, and pP38 protein expression were decreased (P = 0.014, P = 0.004, and P = 0.007, respectively), whereas pAkt expression was induced (P = 0.032).The observed decline of endogenous caveolin-1 protein expression during sepsis implies its involvement in host's cytoprotective reaction either directly, by controlling caveolae population to decrease bacterial burden, or indirectly via regulating 14-3-3b-dependent apoptosis and EPCR-PAR-1-dependent protective signaling.


Subject(s)
Apoptosis , Caveolin 1/biosynthesis , Cytoprotection , Down-Regulation , Lung/metabolism , Sepsis/metabolism , Signal Transduction , Animals , Disease Models, Animal , Lung/pathology , Protein C/pharmacology , Rats , Rats, Wistar , Sepsis/pathology
12.
Mol Med Rep ; 17(2): 2089-2099, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29207125

ABSTRACT

Classic cardiac surgery, determined through the function of cardiopulmonary bypass machine and myocardial cardioplegic arrest, represents the most controlled scenario for cardiomyocyte homeostatic disturbances due to systemic inflammatory response and myocardial reperfusion injury. An increasing number of studies have demonstrated that myocardial cell homeostasis in cardiac surgery procedures is a sequence of molecularly interrelated and overlapping mechanisms in the form of apoptosis, autophagy and necrosis, which are activated by a plethora of induced inflammatory mediators and gene­related signaling pathways. In this study, we outline the molecular mechanisms of the cardiomyocyte adaptive homeostatic process and the associated clinical implications, in the settings of classic cardiac surgery procedures.


Subject(s)
Cardiac Surgical Procedures , Heart/physiology , Homeostasis , Myocardium/metabolism , Animals , Apoptosis , Autophagy , Humans , Myocytes, Cardiac/metabolism , Necrosis
13.
Shock ; 45(2): 139-47, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26513702

ABSTRACT

The impact of a potential autophagy (LC3a/b) deregulation in hyper and in hypo stages during sepsis-induced kidney injury and the temporal profile of phosphorylated extracellular signal-related kinase, P38 (pP38), Akt (pAKT), and 13-3-3ß protein were investigated in the current study, using a rat cecal ligation and puncture (CLP) model, by means of flow cytometry and immunohistochemistry. Cell viability was assessed by protein C zymogen concentrate (PC), 7-aminoactinomycin D (7-AAD) staining and inflammation by S100 protein immunostaining. The impact of reduced kidney inflammation in autophagy was assessed by PC administration, an anti-inflammatory and cytoprotective substance. Sepsis induction increased LC3a/b expression, which presented two peaks at 6 and 36 h after CLP, both in the percentage of positive cells (P = 0.024, P = 0.025, respectively) and in fluorescence intensity. At 6 h when inflammation was already apparent, LC3a/b increase was escorted by phosphorylated extracellular signal-related kinase stimulation and high cell viability (65%), designating autophagy as a cytoprotective mechanism against microbial infection. The phosphorylation of P38 was delayed to 12 h after CLP, when autophagy was reduced. pAkt and 14-3-3ß expression was stimulated between 6 and 36 h after CLP, although a slight inhibition of pAkt within each cell was detected (lower MnIX value). During the second peak, inflammation was intensified, necrosis was significantly increased with LC3a/b+/7-AAD + cells to present a 1.5-fold increase. Protein C zymogen concentrate administration declined autophagy at 6 and 36 h after CLP and reduced necrosis, whereas double positive LC3a/b and 7-AAD cells were increased by 1.68 and 2.78-fold, respectively. These data open new prospectives in sepsis treatment, since they further support that autophagy represents a cytoprotective mechanism triggered by stress conditions, rather than an alternative cell death pathway.


Subject(s)
Acute Kidney Injury/physiopathology , Autophagy/physiology , Inflammation/physiopathology , Sepsis/physiopathology , Animals , Apoptosis/physiology , Cecum/physiopathology , Cells, Cultured , Dactinomycin/analogs & derivatives , Dactinomycin/metabolism , Flow Cytometry , Immunohistochemistry , Ligation , Male , Models, Theoretical , Rats , Rats, Wistar , Signal Transduction/physiology
14.
Int Surg ; 100(4): 696-701, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25875553

ABSTRACT

Laparoscopic surgery results in decreased immune and metabolic stress response compared to open surgery. Our aim was to evaluate the suspension of host immune defense in terms of apoptosis, necrosis, and survival of peripheral T-lymphocytes in patients undergoing laparoscopic versus open cholecystectomy. Apoptosis, necrosis and viability of peripheral T-lymphocytes were measured preoperatively and postoperatively by means of flow cytometry in 27 patients undergoing laparoscopic cholecystectomy and 25 undergoing open cholecystectomy. White cell count, CRP, and serum glucose levels were also measured. Viable peripheral T-lymphocytes were significantly decreased in open cholecystectomy (P = 0.02), while their late apoptotic as well as the overall necrotic rate were significantly increased (P = 0.01 and P < 0.01, respectively). Open cholecystectomy was also associated with lower levels of surviving circulating T-lymphocytes (P = 0.01) and higher percentage of necrotic T lymphocytes (P = 0.03) 24 hours postoperatively compared to laparoscopic cholecystectomy. Serum CRP was increased 24 hours after open cholecystectomy (P = 0.04). All differences failed to sustain more than 48 hours postoperatively. Increased viability and decreased necrosis of circulating T-lymphocytes were observed in laparoscopic cholecystectomy. Necrosis (and not apoptosis) seems to be the predominant pathway of T-lymphocyte death in open cholecystectomy, in a process reaching its peak at 24 hours and further attenuating 48 hours postoperatively.


Subject(s)
Cholecystectomy/methods , T-Lymphocytes/immunology , T-Lymphocytes/pathology , Apoptosis , Blood Glucose/analysis , C-Reactive Protein/analysis , Cholecystectomy, Laparoscopic , Female , Flow Cytometry , Humans , Lymphocyte Count , Male , Middle Aged , Necrosis
15.
Brain Res ; 1587: 1-14, 2014 Oct 31.
Article in English | MEDLINE | ID: mdl-25218612

ABSTRACT

Primary mixed neuronal-astrocytic cultures were established from human brain tissues from elective surgical procedures and maintained in vitro for over 21 days. The majority of cells (a) expressed morphological and cytoskeletal markers of differentiated neurons (MAP2a&b; Tau) or astrocytes (GFAP) in anticipated proportion (1:2), and (b) regenerated synaptic connections and neural-astrocytic associations. Co-cultures with autologous blood leukocytes established that alterations in the viability (by Annexin V/PI) of brain and immune cells over 3 days were indicative of neurodegenerative or immunosuppressive processes. During co-culture, B-cells (CD19+) remained largely unaffected while T-lymphocytes (CD3+) and monocytes (CD14+) declined, consistent with immunosuppressive process. Indications of immunosuppression were not observed when immune cells were maintained in free of neural cells medium collected from neuro-cultures. Decline in brain cell viability in neuro-immune co-cultures may be associated with density of activated monocytes (HLA-DR+/CD14+), consistent with neurodegenerative process. Our findings, though preliminary and associated with significant variability between individuals, establish an approach to investigate neuroimmune pathology in humans.


Subject(s)
Astrocytes/cytology , Leukocytes/cytology , Nerve Degeneration/pathology , Neuroimmunomodulation/physiology , Neurons/cytology , Adult , Antigens, CD/analysis , Astrocytes/metabolism , B-Lymphocytes/cytology , B-Lymphocytes/drug effects , Brain/cytology , Cadherins/biosynthesis , Cadherins/genetics , Cell Communication , Cell Survival , Coculture Techniques , Culture Media, Conditioned/pharmacology , Cytoskeleton/ultrastructure , Gene Expression Profiling , Humans , Immune Tolerance , In Vitro Techniques , Inflammation/pathology , Leukocytes/drug effects , Monocytes/cytology , Monocytes/drug effects , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/genetics , Neurons/metabolism , Primary Cell Culture , T-Lymphocytes/cytology , T-Lymphocytes/drug effects , Wounds and Injuries/blood , Wounds and Injuries/immunology
16.
Clin Exp Metastasis ; 31(5): 511-20, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24590865

ABSTRACT

DNA methylation is the best characterised epigenetic change so far. However, its role in breast cancer metastasis has not as yet been elucidated. The aim of this study was to investigate the differences between the methylation profiles characterising primary tumours and their corresponding positive or negative for metastasis lymph nodes (LN) and correlate these with tumour metastatic potential. Methylation signatures of Caveolin-1, CXCR4, RAR-ß, Cyclin D2 and Twist gene promoters were studied in 30 breast cancer primary lesions and their corresponding metastasis-free and tumour-infiltrated LN with Methylation-Specific PCR. CXCR4 and Caveolin-1 expression was further studied by immunohistochemistry. Tumours were typified by methylation of RAR-ß and hypermethylation of Cyclin-D2 and Twist gene promoters. Tumour patterns were highly conserved in tumour-infiltrated LN. CXCR4 and Caveolin-1 promoter methylation patterns differentiated between node-negative and metastatic tumours. Nodal metastasis was associated with tumour and lymph node profiles of extended methylation of Caveolin-1 and lack of CXCR4 hypermethylation. Immunodetection studies verified CXCR4 and Caveolin-1 hypermethylation as gene silencing mechanism. Absence of Caveolin-1 expression in stromal cells associated with tumour aggressiveness while strong Caveolin-1 expression in tumour cells correlated with decreased 7-year disease-free survival. Methylation-mediated activation of CXCR4 and inactivation of Caveolin-1 was linked with nodal metastasis while intratumoral Caveolin-1 expression heterogeneity correlated with disease progression. This evidence contributes to the better understanding and, thereby, therapeutic management of breast cancer metastasis process.


Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/pathology , Caveolin 1/genetics , Lymph Nodes/pathology , Receptors, CXCR4/genetics , Adult , Aged , Breast Neoplasms/metabolism , Breast Neoplasms/mortality , Caveolin 1/metabolism , DNA Methylation , Female , Gene Expression Profiling , Humans , Immunophenotyping , Lymph Nodes/metabolism , Lymphatic Metastasis , Middle Aged , Neoplasm Grading , Neoplasm Staging , Promoter Regions, Genetic , Receptors, CXCR4/metabolism , Risk Factors
17.
Pathol Res Pract ; 210(2): 105-10, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24268498

ABSTRACT

The expression profiles of 14-3-3ß and θ isoforms, known to exert both oncogenic and antiapoptotic effects, were assessed in different entities of nasal pathophysiology. Flow cytometry and immunohistochemistry were used on paraffin-embedded sections of 51 inverted papillomas (IP), 26 nasal polyps (NP), 9 polyps with IP (NPIP) and 10 specimens of normal epithelium (NE). 14-3-3ß expression was significantly upregulated in IP as compared with both NP (p=0.015) and NE (p=0.002). 14-3-3ß was also increased in NPIP as compared with NE (p=0.008). 14-3-3ß cytoplasmic staining was more pronounced in basal cells of the respiratory epithelium although serous glands and the vascular system were often positive as well. High 14-3-3ß immunopositivity in IP patients concurred with increased proliferative activity shown by PCNA immunostaining (p=0.04). Expression of 14-3-3θ was also found increased in IP and NPIP patients, compared to NP (p=0.005, p=0.002 respectively) and NE (p=0.004 and p=0.001 respectively). 14-3-3θ cytoplasmic immunopositivity was detected in columnar epithelium, particularly in basal and subluminal cells, whereas no immunoreactivity was observed in NP and NE. Our results demonstrate differential expression of 14-3-3ß and θ isoforms in sinonasal pathophysiology, supporting their implication, respectively, in the proliferative and inflammatory process engaged in the formation of IP.


Subject(s)
14-3-3 Proteins/metabolism , Gene Expression Regulation, Neoplastic , Nasal Polyps/metabolism , Nose Neoplasms/metabolism , Papilloma, Inverted/metabolism , Papilloma/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Apoptosis , Cell Proliferation , Cohort Studies , Epithelial Cells , Female , Humans , Male , Middle Aged , Nasal Mucosa/metabolism , Nasal Polyps/physiopathology , Nose Neoplasms/physiopathology , Papilloma/physiopathology , Papilloma, Inverted/physiopathology , Protein Isoforms , Young Adult
18.
Pathol Res Pract ; 208(6): 338-43, 2012 Jun 15.
Article in English | MEDLINE | ID: mdl-22560505

ABSTRACT

Inverted papilloma (IP) is a rare sinonasal benign lesion characterized by aggressive biological behavior. Our aim was to evaluate the expression of various proliferation and apoptotic markers and the presence of HPV genotypes in paraffin sections gathered from surgically treated IP patients. Immunohistochemistry for PCNA, bax, cytochrome c and caspase-8 and flow cytometry for the detection of apoptosis, necrosis and ki67 expression were performed. The identification of various HPV subtypes was achieved by nested PCR amplification. Nasal polyps (NP) and specimens from normal nasal epithelium (NE) were used as controls. PCNA was more frequently expressed in IP compared to NE (p=0.04) and caspase-8 and bax staining were less frequently observed in IP compared to NP (p=0.004 and p=0.01 respectively) and NE (p=0.003 and p=0.01, respectively). IP and NP presented significantly higher Ki67 flow cytometry values compared to NE (p<0.001 and p=0.02 respectively). Cytochrome c was more frequently expressed in IP specimens with more prominent inflammation (p=0.02). A low HPV DNA detection rate was observed. Neither HPV status nor any of the apoptotic or proliferative markers studied was associated with the patients' clinicopathological characteristics. Increased Ki67 appeared to correlate with disease recurrence (p=0.01). Increased PCNA and Ki67 and decreased bax and caspase-8 expression indicate that cell proliferation is increased while apoptosis is inhibited in IP, explaining its biological behavior.


Subject(s)
Nose Neoplasms/pathology , Papilloma, Inverted/pathology , Papillomaviridae/isolation & purification , Tumor Virus Infections/pathology , Adult , Aged , Aged, 80 and over , Apoptosis , Base Sequence , Biomarkers, Tumor/metabolism , Cell Proliferation , DNA, Viral/analysis , Female , Flow Cytometry , Genotype , Humans , Male , Middle Aged , Molecular Sequence Data , Neoplasm Recurrence, Local , Nose Neoplasms/surgery , Nose Neoplasms/virology , Papilloma, Inverted/surgery , Papilloma, Inverted/virology , Papillomaviridae/genetics , Paranasal Sinuses/pathology , Paranasal Sinuses/virology , Proliferating Cell Nuclear Antigen/metabolism , Retrospective Studies , Tumor Virus Infections/complications
19.
J Neurosci Res ; 89(9): 1409-18, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21618583

ABSTRACT

The 14-3-3 family members play a crucial role in the determination of cell fate, exerting their antiapoptotic activity through directly interfering with the critical function of the mitochondrial core proapoptotic machinery. Dimerization of 14-3-3 is vital for the interaction with many of its client proteins and is regulated by phosphorylation. In a previous study, we observed time-dependent neuronal apoptosis during sepsis. Therefore, in the present study, we sought to evaluate the expression of 14-3-3 θ and ß isoforms in septic brain and their association with apoptosis. Sepsis was induced by a CLP model in Wistar rats that were sacrificed at predefined time points. Flow cytometric analysis showed a sepsis-induced, time-dependent alteration of 14-3-3 θ and ß isoforms in both Neun(+) and GFAP(+) cells. 14-3-3 θ was linearly correlated with apoptosis, and stratified analysis for alive and apoptotic neuronal cells demonstrated a gradual down-regulation of θ isoform in alive neurons and astrocytes. The phospho-P38 (pP38) MAP kinase levels were altered in a time-dependent manner during sepsis, presenting a peak at 6 hr post-CLP. A significant correlation between the two isoforms of 14-3-3 was observed in septic rats, with the θ isoform predominant at all time points. The hippocampus, Purkinje cells, and glia-like cells showed intense immunohistochemical reactivity for 14-3-3 θ isoform, whereas the choroid plexus showed constantly increased ß isoform expression. Our results showed that sepsis alters the expression of both 14-3-3 θ and ß isoforms in a time-, cell-, and topography-dependent manner.


Subject(s)
14-3-3 Proteins/metabolism , Apoptosis/physiology , Brain/metabolism , Neurons/metabolism , Sepsis/metabolism , Animals , Brain/pathology , Cecum/surgery , Disease Models, Animal , Male , Neurons/pathology , Protein Isoforms , Rats , Rats, Wistar , Time Factors , Tissue Distribution
20.
J Clin Anesth ; 21(6): 414-21, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19833274

ABSTRACT

STUDY OBJECTIVE: To examine the influence of abdominal colectomy with combined general anesthesia and epidural analgesia versus general anesthesia on apoptosis of circulating lymphocytes. DESIGN: Prospective, randomized, clinical comparison study. SETTING: Tertiary-care general hospital. PATIENTS: 40 ASA physical status I and II patients undergoing elective open colectomy for nonmetastatic colon carcinoma. INTERVENTIONS: Patients were randomly allocated to two groups to receiver either general anesthesia alone (Group G) or general anesthesia combined with epidural analgesia (Group C). Group C comprised 21 patients while 19 patients constituted Group G. All patients underwent median longitudinal laparotomy. MEASUREMENTS: Blood samples were collected preoperatively and 24 hours postoperatively for measurement of lymphocyte apoptosis, serum cortisol, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR). MAIN RESULTS: There were no differences between the two groups in age, weight, or duration of surgery. No significant alterations in total lymphocyte counts, as well as in lymphocyte subpopulations (early apoptotic, late apoptotic, viable, and necrotic), were observed between the general and combined anesthesia groups. Cortisol, ESR, and CRP were significantly increased postoperatively in both groups. Group C presented with lower serum cortisol levels postoperatively than Group G (b = -5.38, CI95%: -8.72 to -2.05, P = 0.002). CONCLUSIONS: Epidural block could not suppress postoperative lymphocyte apoptosis, increases in cortisol, CRP, or ESR compared with general anesthesia.


Subject(s)
Analgesia, Epidural/methods , Anesthesia, General/methods , Apoptosis/drug effects , Colectomy/methods , Aged , Blood Sedimentation/drug effects , C-Reactive Protein/drug effects , C-Reactive Protein/metabolism , Colonic Neoplasms/surgery , Female , Humans , Hydrocortisone/blood , Lymphocytes/drug effects , Lymphocytes/metabolism , Male , Middle Aged , Prospective Studies
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