Sini gene in survival and proliferation of hodgkin and non Hodgkin lymphomas in human

Background: The novel mammalian stress activated protein kinase (SAPK) interacting protein 1 (Sin1) gene give rise to multiple isoforms through alternative splicing. Recent studies provide evidence that Sin1 is a key component of the mammalian target of rapamycin (mTOR)-Rictor complex (mTORC2), thus critically regulating the Ser473 and Thr450-phosphorylation / activation of Akt kinase that results in tumor cell survival through multiple downstream targets. Moreover, in the absence of Sin1 gene, mTOR-Raptor (mTORC1) activity is upregulated upon stimulation of the PI3K/AKT/mTOR pathway. Previous studies have shown that the AKT/mTOR oncogenic pathway is activated in many cancers including aggressive non-Hodgkin lymphomas (NHL). However, the potential role of Sin1 protein in the pathogenesis of B-cell NHL is yet unknown.Methods: The expression of Sin1 gene products was assessed by Western Blot analysis in 26 NHL and HL cell lines and 27 fresh-frozen specimens of NHL of various histologic types. In addition, expression and subcellular localization of Sin1 protein was evaluated using immunohistochemical methods, three tissue microarrays (TMA) of duplicate tumor cores from 81 specimens and 37 samples of paraffin embedded tissues, obtained prior to treatment, which included Reactive Lymph nodes (RL), Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (CLL/SLL), Peripheral T cell Lymphoma Non Otherwise Specified (PTLNOS), Mantle Cell Lymphoma (MCL), Follicular Lymphoma (FL), Marginal Zone Lymphoma (MZL), Diffuse Large B-cell Lymphoma (DLBCL), Anaplastic Large Cell Lymphoma (ALCL), Burkitt Lymphoma (BL) and Multiple Myeloma (MM).Results: Sin1 isoforms 80, 76, 55 and 52 kDa were differentially expressed in immunoblots of B and T-cell NHL tested. Sin1 protein levels and activation status of its downstream target Akt, were correlated with the histological grade. This association was also seen in primary NHL tumor specimens analyzed by Western blot. Immunohistochemical analysis of Sin1 protein revealed that Sin1 was localized in the cytoplasm of tumor cells in low grade NHL, MCL and MM and in the nucleus in high grade NHL (p<0.001, chi-square test). In addition strong nuclear expression of mSin1 in high grade B-NHL and cytoplasmic in low grade B-NHL (p<0.001, chi-square test) was observed. Transfection of MSCV-Sin1.1 and MSCV-Sin1.5 plasmids to SP53 cells resulted in increased cell viability and cell growth, which was associated with decreased apoptosis. These biologic effects were associated with upregulation of Bcl-2 and Bcl-XL as well as c-FLIP suggesting downregulation of both mitochondrial and extrinsic apoptotic pathways, respectively. Conclusions: Sin1, a critical regulator of mTORC2 integrity and activity, is differentially expressed among B-NHL. Sin1 expression seems to confer AKT-dependent anti-apoptotic signals in MCL cells and possibly in other in vitro systems of B-NHL.Εισαγωγή: Το γονίδιο που κωδικοποιεί τη πρωτεΐνη που αλληλεπιδρά με τις επαγόμενες από το στρες πρωτεϊνικές κινάσες mSin1 (Stress activated protein kinase INteracting protein 1 Sin1) με εναλλακτική συρραφή σχηματίζει πέντε διαφορετικές ισομορφές. Πρόσφατες μελέτες υποδεικνύουν ότι η mSin1 αποτελεί βασικό συστατικό του συμπλόκου 2 της κινάσης στόχου της ραπαμυκίνης (Target Of Rapamycin TOR) των θηλαστικών (mTORC2) και ρυθμίζει την ενεργοποίηση της Akt, με φωσφορυλίωση στην Ser473 και την Thr450, που επάγει την επιβίωση μέσω πολλαπλών καθοδικών στόχων. Απουσία της mSin1 η ενεργότητα του συμπλόκου mTORC1 αυξάνεται επαγόμενη από την σηματοδοτική οδό PI3K/Akt/mTOR. Έχει δειχτεί ότι η ογκογόνος οδός Akt/mTOR είναι ενεργοποιημένη σε πολλούς τύπους καρκίνου και σε επιθετικά μη- Hodgkin λεμφώματα (NHL). Ωστόσο ο ρόλος της mSin1 στην παθογένεια των Β μη- Hodgkin λεμφωμάτων δεν είναι γνωστός.Μέθοδοι: Η έκφραση της mSin1 μελετήθηκε με Western Blot σε 26 NHL και HL κυτταρικές σειρές και πρωτογενή κύτταρα και 27 φρέσκους ιστούς NHL διαφόρων ιστολογικών τύπων. Επίσης η έκφραση και ο υποκυτταρικός εντοπισμός της mSin1 αξιολογήθηκε με ανοσοϊστοχημεία σε ιστούς εγκλεισμένους σε παραφίνη πριν την χορήγηση θεραπείας σε 118 ασθενείς με αντιδραστική λεμφαδενίτιδα, λέμφωμα από μικρά κύτταρα/χρόνια λεμφογενής λευχαιμία, λέμφωμα του μανδύα, οζώδες λέμφωμα, ,λέμφωμα οριακής ζώνης, διάχυτο λέμφωμα από μεγάλα κύτταρα, αναπλαστικό λέμφωμα από μεγάλα κύτταρα, λέμφωμα Burkitt, πολλαπλό μυέλωμα, περιφερικό Τ λέμφωμα μη-προσδιοριζόμενο. Αποτελέσματα: Οι ισομορφές με μέγεθος 80, 76, 55 και 52 kDa παρουσίασαν διαφορική έκφραση στα NHL που εξετάστηκαν. Τα επίπεδα της Sin1 και η ενεργοποίηση της Akt, συσχετίζονταν με τον ιστολογικό βαθμό κακοήθειας. Η συσχέτιση αυτή παρατηρήθηκε και σε πρωτοπαθή NHL δείγματα που μελετήθηκαν με Western blot. Η ανοσοϊστοχημική ανάλυση αποκάλυψε στατιστικά σημαντική διαφορά στον υποκυτταρικό εντοπισμό της Sin1 στα υψηλής και χαμηλής κακοήθειας Β λεμφώματα, το ΜΜ και το MCL (p<0.001, chi-square test). Στατιστικά σημαντική ήταν και η έντόπιση της Sin1 ανάμεσα στα υψηλής και χαμηλής κακοηθείας Β λεμφώματα (p<0.001, chi-square test). Παροδική διαμόλυνση κυττάρων SP53 με πλασμίδια MSCV-Sin1.1 και MSCV-Sin1.5 επέφερε αύξηση του συνολικού αριθμού των κυττάρων και της επιβίωσης, σε συνάρτηση με μείωση της απόπτωσης. Αυτό το βιολογικό αποτέλεσμα συσχετιζόταν με μείωση των of Bcl-2 και Bcl-XL, καθώς και του c-FLIP, υποδεικνύοντας αρνητική ρύθμιση της ενδογενούς και εξωγενούς αποπτωτικού μηχανισμού. Συμπεράσματα: Η Sin1 αποτελεί κρίσιμο ρυθμιστή της συγκρότησης και ενεργότητας του συμπλόκου mTORC2, διαφορικά εκφραζόμενη στα B-NHL. Η έκφραση της Sin1 συμβάλλει στους εξαρτώμενους από την Akt αντιπαποπτωτικούς μηχανισμούς στο MCL και πιθανόν και σε άλλα in vitro συστήματα B-NHL

DNA repair systems in rhabdomyosarcoma.

Rhabdomyosarcoma (RMS) represents the most common soft tissue sarcoma in children and adolescent population. There are two major histological subtypes, embryonal (ERMS) and alveolar (ARMS), differing in cytogenetic and morphological features. RMS pathogenesis remains controversial and several cellular mechanisms and pathways have been implicated. Application of intense chemo- and radio-therapy improves survival rates for RMS patients, but significant efficacy has not been proved as DNA damage induced-resistance frequently occurs. The present review is aimed at summarizing the current evidence on DNA repair systems, implications in RMS development, focusing on gene expression alterations and point mutations of genes encoding for DNA repair enzymes. Understanding of DNA repair systems involvement in RMS pathogenesis could diversify RMS patients and provide novel individualized therapeutic targets

DNA repair systems in rhabdomyosarcoma

Rhabdomyosarcoma (RMS) represents the most common soft tissue sarcoma in children and adolescent population. There are two major histological subtypes, embryonal (ERMS) and alveolar (ARMS), differing in cytogenetic and morphological features. RMS pathogenesis remains controversial and several cellular mechanisms and pathways have been implicated. Application of intense chemo- and radiotherapy improves survival rates for RMS patients, but significant efficacy has not been proved as DNA damage induced-resistance frequently occurs. The present review is aimed at summarizing the current evidence on DNA repair systems, implications in RMS development, focusing on gene expression alterations and point mutations of genes encoding for DNA repair enzymes. Understanding of DNA repair systems involvement in RMS pathogenesis could diversify RMS patients and provide novel individualized therapeutic targets

Carotid Disease and Ageing: A Literature Review on the Pathogenesis of Vascular Senescence in Older Subjects

Aging is a natural process that affects all systems of the human organism, leading to its inability to adapt to environmental changes. Advancing age has been correlated with various pathological conditions, especially cardiovascular and cerebrovascular diseases. Carotid artery (CA) is mainly affected by age-induced functional and morphological alterations causing atheromatous disease. The evolvement of biomedical sciences has allowed the elucidation of many aspects of this condition. Symptomatic carotid disease (CD) derives from critical luminar stenosis or eruption of an atheromatous plaque due to structural modifications of the vessels, such as carotid intima-media thickening. At a histologic level, the aforementioned changes are mediated by elastin fragmentation, collagen deposition, immune cell infiltration, and accumulation of cytokines and vasoconstrictors. Underlying mechanisms include chronic inflammation and oxidative stress, dysregulation of cellular homeostatic systems, and senescence. Thus, there is an imbalance in components of the vessel wall, which fails to counteract exterior stress stimuli. Consequently, arterial relaxation is impaired and atherosclerotic lesions progress. This is a review of current evidence regarding the relationship of aging with vascular senescence and CD. A deeper understanding of these mechanisms can contribute to the production of efficient prevention methods and targeted therapeutic strategies

Comprehensive Molecular Analysis of NSCLC; Clinicopathological Associations.

Selection of NSCLC patients for targeted therapy is currently based upon the presence of sensitizing mutations in EGFR and EML4/ALK translocations. The heterogeneity of molecular alterations in lung cancer has led to the ongoing discovery of potential biomarkers and targets in order to improve survival.This study aimed to detect alterations in EGFR, KRAS, BRAF, PIK3CA, MET-gene copy number and ALK rearrangements in a large cohort of 956 NSCLC patients of Hellenic origin using highly sensitive techniques and correlations with clinicopathological characteristics.Mutations were detected in EGFR 10.6% (101 out of 956 samples), KRAS 26.5% (191 out of 720 samples), BRAF 2.5% (12 out of 471 samples), PIK3CA 3.8% (7 out of 184 samples), MET gene amplification was detected in 18% (31 out of 170) and ALK rearrangements in 3.7% (4 out of 107 samples). EGFR mutations were detected in exon 19 (61.4% of mutant cases), exon 21 p.Leu858Arg (19.8%), exon 20 (15.8%), exon 18 (2.9%) and were correlated with gender histology, smoking status and TTF1 staining. p.Thr790Met mutant cases (3.9%) displayed concurrent mutations in exons 19 or 21. Negative TTF-1 staining showed strong negative predictive value for the presence of EGFR mutations. KRAS mutations were associated with histology, the most common mutation being p.Gly12Cys (38%).In conclusion, only 89 patients were eligible for EGFR -TKIs and ALK inhibitors therapy, whereas 257 patients showed other alterations, highlighting the necessity for a detailed molecular profiling potentially leading to more efficient individualized therapies for NSCLC patients

Comprehensive Molecular Analysis of NSCLC; Clinicopathological Associations

Background Selection of NSCLC patients for targeted therapy is currently based upon the presence of sensitizing mutations in EGFR and EML4/ALK translocations. The heterogeneity of molecular alterations in lung cancer has led to the ongoing discovery of potential biomarkers and targets in order to improve survival. Aim This study aimed to detect alterations in EGFR, KRAS, BRAF, PIK3CA, MET-gene copy number and ALK rearrangements in a large cohort of 956 NSCLC patients of Hellenic origin using highly sensitive techniques and correlations with clinicopathological characteristics. Results Mutations were detected in EGFR 10.6%(101 out of 956 samples), KRAS 26.5% (191 out of 720 samples), BRAF 2.5% (12 out of 471 samples), PIK3CA 3.8% (7 out of 184 samples), MET gene amplification was detected in 18% (31 out of 170) and ALK rearrangements in 3.7% (4 out of 107 samples). EGFR mutations were detected in exon 19 (61.4% of mutant cases), exon 21 p.Leu858Arg (19.8%), exon 20 (15.8%), exon 18 (2.9%) and were correlated with gender histology, smoking status and TTF1 staining. p.Thr790Met mutant cases (3.9%) displayed concurrent mutations in exons 19 or 21. Negative TTF-1 staining showed strong negative predictive value for the presence of EGFR mutations. KRAS mutations were associated with histology, the most common mutation being p.Gly12Cys (38%). Discussion In conclusion, only 89 patients were eligible for EGFR-TKIs and ALK inhibitors therapy, whereas 257 patients showed other alterations, highlighting the necessity for a detailed molecular profiling potentially leading to more efficient individualized therapies for NSCLC patients

Carotid Disease and Ageing: A Literature Review on the Pathogenesis of Vascular Senescence in Older Subjects.

Aging is a natural process that affects all systems of the human organism, leading to its inability to adapt to environmental changes. Advancing age has been correlated with various pathological conditions, especially cardiovascular and cerebrovascular diseases. Carotid artery (CA) is mainly affected by age-induced functional and morphological alterations causing atheromatous disease. The evolvement of biomedical sciences has allowed the elucidation of many aspects of this condition. Symptomatic carotid disease (CD) derives from critical luminar stenosis or eruption of an atheromatous plaque due to structural modifications of the vessels, such as carotid intima-media thickening. At a histologic level, the aforementioned changes are mediated by elastin fragmentation, collagen deposition, immune cell infiltration, and accumulation of cytokines and vasoconstrictors. Underlying mechanisms include chronic inflammation and oxidative stress, dysregulation of cellular homeostatic systems, and senescence. Thus, there is an imbalance in components of the vessel wall, which fails to counteract exterior stress stimuli. Consequently, arterial relaxation is impaired and atherosclerotic lesions progress. This is a review of current evidence regarding the relationship of aging with vascular senescence and CD. A deeper understanding of these mechanisms can contribute to the production of efficient prevention methods and targeted therapeutic strategies

Frequencies of alterations between different groups of adenocarcinomas (NL = Normal, MT = Mutant, AMPL = gene amplification).

<p>Frequencies of alterations between different groups of adenocarcinomas (NL = Normal, MT = Mutant, AMPL = gene amplification).</p

KRAS mutations distributed amongst different Adenocarcinoma groups.

<p>KRAS mutations distributed amongst different Adenocarcinoma groups.</p