Journal of molecular medicine and clinical applications最新文献

{"title":"Identification of appropriate housekeeping genes for quantitative RT-PCR analysis in MDA-MB-231 and NCI-H460 human cancer cell lines under hypoxia and serum deprivation","authors":"A. Albuquerque, M. Balmaña, C. Reis, E. Beltrão","doi":"10.31083/J.JMCM.2018.03.001","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.001","url":null,"abstract":"This work was supported by the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) [grant agreement No. 88881.132780/2016-01]; Fundac ao de Amparoa Ciencia e Tecnologia de Pernambuco (FACEPE) [grant agreement No. 0883-2.08/13];and the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie [grant agreement No. 748880]. This work was also funded by the project NORTE-01-0145-FEDER-000029, supported by Norte Portugal Regional Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and by FEDER—Fundo Europeu de Desenvolvimento Regional funds through COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020,and by Portuguese funds through FCT (Fundacao para a Ciencia e a Tecnologia)/Ministerio da Ciencia, Tecnologia e Inovacao in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274), and the project POCI-01-0145-FEDER-016585 (PTDC/BBB EBI/0567/2014).","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85000574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overcoming palbociclib resistance by combined treatment with PI3K/AKT/mTOR inhibitors in mesothelioma cells","authors":"Graziana Digiacomo, C. Fumarola, Daniele Cretella, R. Alfieri, S. L. Monica, P. Petronini, M. Bonelli, A. Cavazzoni","doi":"10.31083/J.JMCM.2018.03.004","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.004","url":null,"abstract":"Carcinogenesis of malignant pleural mesothelioma (MPM) is strictly associated with chronic exposure of mesothelial cells from the pleura to asbestos fibers and MPM incidence is expected to peak in the next years. Extensive genome analyses of patientderived MPM tumors revealed that the most frequent mutational event involves the inactivation of the CDKN2A gene that encodes for the cell cycle inhibitors p16INK4a and p14ARF with consequent constitutive activation of CDK4/6 – cyclin D complexes. Therefore, inhibition of the latter complexes may represent a new option for the treatment of MPM patients. However, despite the efficacy of the specific CDK4/6 inhibitor palbociclib in blocking MPM cell proliferation, acquired resistance inevitably occurs. Herein, palbocilib-resistant clones isolated after stepwise exposure of MSTO-211H cells to gradually increasing drug concentrations, showed a reduction in Rb protein levels as well as in the cell cycle inhibitor p21waf1, accompanied by increased phosphorylation of AKT and p70S6K. Simultaneous treatment of resistant clones with both palbociclib and specific PI3K/AKT/mTOR inhibitors produced an additive effect in terms of reduction in cell growth, without any signs of senescence but with increased cell death. In MSTO-211H sensitive cells, this combination drug treatment significantly delayed the adaptation to palbociclib, suggesting that this treatment approach may prevent or at least retard the emergence of palbociclib resistance. Collectively, these results suggest that the combination of palbociclib with PI3K/AKT/mTOR inhibitors may overcome the acquisition of resistance to palbociclib treatment and could represent a potential therapeutic approach to treat cancers with acquired resistance to CDK4/6 inhibitors in the presence of activation of the AKT/mTOR signaling pathway.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91104665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Semaphorin-3A levels in urine demonstrates promising sensitivity for detection of upper-tract urothelial carcinoma – a preliminary case series","authors":"Itamar Getzler, Z. Vadasz, J. Rubinstein, S. Halachmi","doi":"10.31083/J.JMCM.2018.03.008","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.008","url":null,"abstract":"The purpose of this study was to evaluate the utility of Semaphorin-3A (Sema-3A) as a biomarker for diagnosis and management of upper tract urothelial carcinoma, independently and in conjunction to cytology. Upper tract noninvasive urothelial carcinoma is a lifelong chronic non-curable disease. It is infamously difficult both to diagnose and to follow-up, and current non-invasive methods are commonly inadequate. Sema-3A protein levels can be measured from a simple urine test and can aid in the diagnosis and follow-up and early non-invasive tumor recurrence detection. Assigned cases for this series were those with pathologically verified upper tract neoplastic lesions. Urine samples for cytology and Sema-3A were taken on admission from all recruited patients. Sema-3A protein levels were determined using ELISA in every sample, and the tumor was graded and staged according to the 2004 WHO grading system. Descriptive and statistical analysis was performed to evaluate the performance of Sema-3A and the cytology exam. This case series included 10 patients with pathologically proven upper tract urothelial carcinoma. Sensitivity for recognizing disease was calculated as 80% for cytology, and 89% for a predetermined Sema-3A cutoff. Combining the strengths of both urine tests to a single criterion (Sema-3A > 5 or positive cytology) resulted in 100% sensitivity. In conclusion, in the current preliminary study, high levels of Sema-3A correlated with upper-tract urothelial cancer stage and displayed higher sensitivity than cytology. Combined analysis of both positive Sema-3A and cytology demonstrated 100% sensitivity. Thus, Sema-3A levels can potentially serve as a reliable biomarker for diagnosis and management for the disease, given that further dedicated studies with larger patient cohorts are undertaken.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78497705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SIRPα is transcriptionally downregulated by epigenetic silencing in medulloblastoma","authors":"D. Vuurden, E. Hulleman, Mahban Irandoust, Dennis Biesmans, T. Berg, E. Aronica, V. Hovestadt, M. Kool, W. Vandertop, G. Kaspers, J. Cloos","doi":"10.31083/J.JMCM.2018.03.005","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.005","url":null,"abstract":"Signal regulatory protein α (SIRPα) is a transmembrane protein that is commonly expressed in cells of the hematopoietic system and brain. Its function is not fully understood but it includes tumor suppressor properties and effects on differentiation. SIRPα may play a role in the development of medulloblastoma (MB), a WHO grade IV brain tumor, which is the most common malignant brain tumor in childhood. The aim of the current study was to determine the possible role of SIRPα in MB cells. Interestingly, in contrast to normal cerebellum, SIRPα mRNA was strongly downregulated in MB and its protein was not detectable in MB tissues. This down-regulation in MB cells was associated with transcriptional silencing of SIRPα via CpG island promoter hypermethylation. Furthermore, Oncomir cluster miR17-92 and miR-106a were correlated with SIRPα gene silencing in MB tumor specimens and cell lines. Histone modification and inhibition of DNA methylation using TSA (20 nM) for 24 hrs and 5-AZA (5 μM) and DZnep (2.5 μM) for 72 hrs, respectively, increased SIRPα expression 25-40 fold and resulted in 90% cytotoxicity of MB tumor cell lines D283-med and D458-med. Remarkably, forced upregulation of SIRPα by viral transduction in MB cell lines did not affect cell growth. In conclusion, SIRPα is epigenetically silenced in MB cells and tumor specimens by promoter hypermethylation and possibly by miRNA expression. SIRPα hypermethylation in MB might reflect the precursor cell state of these cells, rather than being a tumor-specific event, since SIRPα overexpression did not influence MB cell viability. The mechanism of the anti-MB action of epigenetic therapy requires further investigation since our findings indicate that this effect is independent of SIRPα upregulation.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87814666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lateral Guided Bone Regeneration Using a Novel Synthetic Bioresorbable Membrane: A Two Center Prospective Randomized Controlled Trial","authors":"Michal Halperin-Sternfeld, H. Zigdon-Giladi, L. Shapira, A. Wilensky","doi":"10.31083/J.JMCM.2018.03.006","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.006","url":null,"abstract":"The aim of this study was to evaluate the outcomes of lateral guided bone regeneration (GBR) using a novel resorbable synthetic polyethylene-glycol/methacrylate (PEG/MET) membrane compared to a non-cross-linked collagen membrane (CM). Twentyeight patients with a potential implant site exhibiting insufficient bone width of ≤ 5 mm were included. Ridge width was measured intraoperatively at 1 mm and 4 mm apical to the crest and via cone-beam computed tomography at baseline and 6 months following GBR using either a PEG/MET or a CM in conjunction with an allograft. During implant placement, core biopsies were harvested and analyzed histomorphometrically. Width changes were calculated. Differences between groups were analyzed using two-sided t-test and Mann-Whitney U-test. The PEG/MET membrane was moldable and exhibited higher strength and stability compared to the CM. Nevertheless, it displayed higher exposure rate of 12/15, compared to 2/13 in the CM sites. At the time of implant insertion, 6 months following GBR, significant gain in bone width was observed in both groups. Mean ridge width at 1 mm and 4 mm apical to the crest was increased significantly from 2.06 ± 0.77 mm and 3.84 ± 1.23 mm to 3.84 ± 1.52 mm and 6.06 ± 2.03 mm (p = 0.0006 and p = 0.0009, respectively), with no clinical or radiographic differences between groups. Experimental sites contained more residual scaffold material than the controls (17.4 ± 3.3% and 8.6 ± 2.0%, p = 0.0566). However, bone and connective tissue area fraction were not statistically different between the groups. Overall, despite the higher exposure rate, the new PEG/MET membrane was as successful as a standard collagen membrane in lateral GBR and may have potential use in bone augmentation procedures. This study suggests the feasibility of synthetic membranes, which are not associated with disease transmission, as an attractive alternative to the commonly used CM of bovine or porcine origin.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74602378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preoperative neutrophil to lymphocyte ratio can improve disease progression prediction of non-muscle invasive bladder cancer","authors":"Itamar Getzler, O. Nativ, R. Mano, J. Baniel, J. Rubinstein, S. Halachmi","doi":"10.31083/J.JMCM.2018.03.002","DOIUrl":"https://doi.org/10.31083/J.JMCM.2018.03.002","url":null,"abstract":"The purpose of this study was prospectively evaluate the ability of Neutrophil-to-Lymphocyte ratio (NLR) to predict disease progression in patients with non-muscle invasive bladder cancer (NMIBC). This is a continuation of our previous retrospective study that indicated the significance of NLR > 2.5 criterion as a predictor of progression in patients with NMIBC. Since December 2013, all patients admitted to Bnai-Zion department for TUR-BT and agreed to participate in the study, had blood analyses for cell count and differential 24hr prior to surgery. Patients with pathological NMIBC were followed prospectively for disease progression. The end-point of the follow up was either a disease progression or the termination of the study. Kaplan-Meier curves and Cox regression were performed to assess the predictive ability of NLR > 2.5 for disease progression. Our results demonstrate a significant difference (p = 0.02) in mean progression-free survival – (35.9 months vs 41.1 months) in the whole cohort Kaplan-Meier survival plot factored by NLR > 2.5. Mean progression-free survival of NLR > 2.5 stratified by stage, grade and treatment (sub-group analysis), showed statistical significance (p= 0.035) for those treated with intra-vesical instillation, and demonstrated a persistent trend for the rest of the stratifications revealing that the NLR > 2.5 groups always fared worse than the NLR < 2.5 groups. In a univariate analysis, whole cohort Cox regression analysis for disease progression, NLR > 2.5 was found significant (p= 0.05; HR 7.8; CI 1–61), indicating that the probability of progression is increased at least 7-fold for a person with a NLR > 2.5 compared with those with NLR < 2.5. In conclusion, NLR > 2.5 was found to be a significant predictor of disease progression and demonstrated high hazard ratio and worse progression-free survival in patients with NMIBC, especially in those treated with intra-vesical instillation. We propose to consider the incorporation of NLR > 2.5 in the next revisions of the European Organization for Research and Treatment of Cancer (EORTC) scores, given more widely available evidence.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88754654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DPYD gene activity score (GAS) predicts dose-limiting toxicity in fluoropyrimidine-treated colorectal cancer patients","authors":"Dalle Fratte Chiara, P. Jerry, Roncato Rossana, Delmastro Elena, Ecca Fabrizio, Bignucolo Alessia, Garziera Marica, Dreussi Eva, P. Elisa, Buonadonna Angela, G. Michela, B. Massimiliano, Foltran Luisa, Sartor Franca, D. Mario, Favaretto Adolfo, M. Enrico, N. Stefania, D. Antonino, T. Giuseppe, C. Erika","doi":"10.31083/j.jmcm.2018.03.003","DOIUrl":"https://doi.org/10.31083/j.jmcm.2018.03.003","url":null,"abstract":"Pre-treatment DPYD genotyping of a panel of 4-single nucleotide polymorphism (SNP) including DPYD*2A, DPYD*13, c.2846A>T and c.1236A>G-HapB3, has been strongly recommended by the current pharmacogenetics guidelines in order to avoid severe fluoropyrimidine (FL)-related toxicity. However, translation to clinical practice is still lagging behind. This requires a better definition of the relationship between genetic variants of DPYD and dose-limiting toxicities (DLTs) and development of new methods to investigate the effect of DPYD variants, such as the DPYD activity score. The aim of the current study was to support the clinical implementation of DPYD genetic testing, by assessing the relationship between DPYD variants in the 4-SNP panel and the risk to develop DLTs and by stratifying patients according to the DPYD gene activity score (GAS) model. (GAS = 1.0 if carriers of one DPYD*2A or DPYD*13 alleles and GAS = 1.5, if carriers of one c.2846A>T or c.1236G>A-HapB3 allele. Non-carriers GAS = 2.0). A retrospective population of 763 colorectal cancer patients treated with FL-based chemotherapy, was selected and genotyped. Patients carrying at least one decreased function DPYD variant in the 4-SNP panel, displayed a significant association with the risk of developing DLT (i.e. grade ≥ 3 non-hematological toxicity or grade ≥ 4 hematological toxicity) either within the first three cycles of chemotherapy (OR= 2.7, 95% CI = 1.33–5.41) or during the entire course of treatment (OR = 2.7, 95% CI = 1.42–5.04). Patients’ GAS was found to better define the risk of DLT for both acute (GAS = 1.5, OR = 1.80, 95% CI = 0.78–4.15 and GAS = 1.0, OR = 10.12, 95% CI = 2.55–40.20) and total toxicity (GAS = 1.5, OR = 2.08, 95% CI = 1.02–4.27 and GAS = 1, OR = 7.09, 95% CI = 1.69–29.65). In conclusion, the present study demonstrates that the DPYD 4-SNP panel and the associated GAS can predict the occurrence of DLT related to treatment with FL. These findings further support the implementation of pre-emptive DPYD genotyping in the routine clinical practice.","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86764723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Docking: From Lock and Key to Combination Lock.","authors":"Ashutosh Tripathi,&nbsp;Vytas A Bankaitis","doi":"10.16966/2575-0305.106","DOIUrl":"https://doi.org/10.16966/2575-0305.106","url":null,"abstract":"<p><p>Accurate modeling of protein ligand binding is an important step in structure-based drug design, is a useful starting point for finding new lead compounds or drug candidates. The 'Lock and Key' concept of protein-ligand binding has dominated descriptions of these interactions, and has been effectively translated to computational molecular docking approaches. In turn, molecular docking can reveal key elements in protein-ligand interactions-thereby enabling design of potent small molecule inhibitors directed against specific targets. However, accurate predictions of binding pose and energetic remain challenging problems. The last decade has witnessed more sophisticated molecular docking approaches to modeling protein-ligand binding and energetics. However, the complexities that confront accurate modeling of binding phenomena remain formidable. Subtle recognition and discrimination patterns governed by three-dimensional features and microenvironments of the active site play vital roles in consolidating the key intermolecular interactions that mediates ligand binding. Herein, we briefly review contemporary approaches and suggest that future approaches treat protein-ligand docking problems in the context of a 'combination lock' system.</p>","PeriodicalId":92248,"journal":{"name":"Journal of molecular medicine and clinical applications","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.16966/2575-0305.106","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35736546","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}