Stress-Related Regulation Is Abnormal in the Psoriatic Uninvolved Skin

Keratinocyte stress-response of the uninvolved psoriatic epidermis is known to be altered compared to healthy cells. Therefore, we aimed to reveal potential mechanisms underlying this alteration. We compared the expression of annotated cell-stress-related proteins between uninvolved psoriatic and healthy skin using the protein array method. Data were analyzed by the Reactome over-representation test. We found that p27/CDKN1B and cytochrome C showed at least a two-fold increase, while cyclooxygenase-2, indolamine-2,3-dioxygenase-1, serum paraoxonase 1, serum paraoxonase 3, serine-46-phosphorylated tumor protein p53, and superoxide-dismutase-2 showed a two-fold decrease in expression in the uninvolved skin. Over-representation analysis suggested the Forkhead-box protein O (FOXO)-mediated transcription as the most significant pathway affected by the differently expressed cell-stress-related proteins (DECSRPs). DECSRPs indicate increased FOXO-mediated transcription of cell-cycle genes and reduced interleukin-signaling in the psoriatic uninvolved skin. Nuclear positivity of the FOXO-signaling-related p27/CDKN1B and FOXO1 are negatively correlated with the disease severity and showed increased expression in the uninvolved epidermis and also in healthy primary keratinocytes, which were grown on cartilage oligomeric matrix protein-coated surfaces. Our results indicate a cell-cycle inhibitory process, as a stress-related compensatory mechanism in the uninvolved epidermis, that could be responsible for blocking keratinocyte hyperproliferation in the psoriatic uninvolved skin, thus maintaining the symptomless skin phenotype.

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FMNL1 Is Downregulated In Myelodysplastic Syndromes and Is Involved In Megakaryocytic Differentiation

Blood ◽

10.1182/blood.v122.21.1519.1519 ◽

2013 ◽

Vol 122 (21) ◽

pp. 1519-1519

Author(s):

Matheus Rodrigues Lopes ◽

João Agostinho Machado-Neto ◽

Fabiola Traina ◽

João Kleber Novais Pereira ◽

Irene Lorand-Metze ◽

...

Keyword(s):

Cell Cycle ◽

Bone Marrow ◽

Bone Marrow Cells ◽

K562 Cells ◽

Fold Increase ◽

Granulocytic Differentiation ◽

Megakaryocytic Differentiation ◽

Total Bone Marrow ◽

Related Proteins ◽

Dependent Processes

Abstract Background Myelodysplastic syndromes (MDS) are disorders characterized by morphological dysplasia, impaired differentiation and defective cellular functions, resulting in peripheral cytopenias. FMNL1 belongs to a family of formin-related proteins, indispensable for many fundamental actin-dependent processes. Recently, FMNL1 has been described to be upregulated and play a role in the actin cytoskeleton dynamics during monocyte differentiation to macrophages. Aims The aim of this work was to characterize FMNL1 expression in total bone marrow cells of patients with MDS comparing to normal donors. We also analyzed FMNL1 expression in erythrocytic, granulocytic and megakaryocytic differentiation, using cell line models. Finally, we evaluated the impact of inhibition of FMNL1 during megakaryocytic differentiation. Methods A total of 49 patients with a diagnosis of MDS, receiving no treatment, and 18 samples from normal donors were included in the study, which was approved by the National Ethical Committee Board. Samples were submitted to RNA extraction after removal of erythrocytes by hemolysis. FMNL1 expression levels from cell lines or total bone marrow cells were determined by quantitative PCR (q-PCR) or Western blot. KU812 was treated 50 μM hemin and 100 μM hydroxyurea for erythrocytic differentiation. K562 was stimulated with 20nM of PMA for megakaryocytic differentiation. NB4 was treated with 10-6 M of ATRA for granulocytic differentiation. Megakaryocytic differentiation was followed by the increase in megakaryocytic marker (CD61) determined by flow cytometry and cells were also stained with May–Grunwald–Giemsa. K562 cells were transduced with lentivirus-mediated shRNA targeting LacZ or FMNL1. Apoptosis was assessed by Annexin-V/PI staining and cell cycle was evaluated by flow cytometry, both at 24 and 48 hours after induction with PHA. The statistical methods used were the age-adjusted multivariate linear regression analysis, Mann Whitney test or t test. Results FMNL1 expression in bone marrow samples was significantly lower in MDS when compared with normal donor cells (P=0.01), especially in the high risk group (P<0.02). Using cell line models for hematopoietic differentiation, there was a fifteen-fold increase and a five-fold increase in FMNL1 expression for megakaryocytic (P=0.002) and granulocytic differentiation (P=0.05) respectively. Western blot analysis corroborated these findings. There was no difference for erythrocytic differentiation. After PMA treatment, the level of the megakaryocytic markers CD61 was significantly lower in K562 shFMNL1 when compared with shLacZ (P=0.01). The level of CD41a (P=0.5) and CD42b (P=0.1) showed a trend toward a decrease in K562 shFMNL1 when compared with shLacZ, however not statistically significantly. Corroborating these data, K562 shFMNL1 showed abnormal megakaryocytic morphological features (larger cells with polylobulated or polysegmented nuclei and vacuolization) compared with cells shLacZ. There were no statistical differences in the apoptosis levels and cell cycle analysis between K562 shFMNL1 and shLacz cells. Conclusions The family of formin-related proteins has mainly been related to actin-dependent processes although little is known regarding their possible involvement in haematopoiesis. The lower FMNL1 expression in MDS BM could reflect the role of this protein in cell differentiation. We were therefore prompted to study this issue in depth using megakaryocytic differentiation as a system. In this study, we show increased expression of FMNL1 in PMA-induced megakaryocytic differentiation of K562 cells. Furthermore, knockdown of FMNL1 deregulates differentiation, suggesting that FMNL1 is required in order to maintain the effective megakaryocytopoiesis in MDS. Although FMNL1 silencing effectively down-regulated CD61 expression, CD41a and CD42b were reduced to a lesser extent. The effect of FMNL1 cannot be explained by modifications of the cell cycle or apoptosis during differentiation; and is probably due to the effect in changes in the dynamic remodeling of the cytoskeleton. Disclosures: No relevant conflicts of interest to declare.

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Influence of AUF1 targeting siRNA on cell cycle-related proteins in thyroid cancer cell lines

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-2006-932951 ◽

2006 ◽

Vol 114 (S 1) ◽

Author(s):

B Trojanowicz ◽

Z Chen ◽

J Bialek ◽

Y Radestock ◽

S Hombach-Klonisch ◽

...

Keyword(s):

Cell Cycle ◽

Thyroid Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Thyroid Cancer Cell ◽

Thyroid Cancer Cell Lines ◽

Related Proteins

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Faculty Opinions recommendation of Catalytic versatility and backups in enzyme active sites: the case of serum paraoxonase 1.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.723676156.793514594 ◽

2016 ◽

Author(s):

Jeffrey Skolnick

Keyword(s):

Active Sites ◽

Paraoxonase 1 ◽

Enzyme Active Sites ◽

Serum Paraoxonase

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Deregulation of cell cycle and related microRNA expression induced by vinyl chloride monomer in hepatocytes of rats

Toxicology and Industrial Health ◽

10.1177/07482337211015591 ◽

2021 ◽

pp. 074823372110155

Author(s):

Weizhe Pan ◽

Shengnan Yu ◽

Jin Jia ◽

Junyang Hu ◽

Liang Jie ◽

...

Keyword(s):

Cell Cycle ◽

Vinyl Chloride ◽

Male Rats ◽

Cell Cycle Distribution ◽

Vinyl Chloride Monomer ◽

Dynamic Changes ◽

Cell Cycle Deregulation ◽

Change Dynamic ◽

Related Proteins

Vinyl chloride (VC) is a confirmed human carcinogen associated with hepatocellular carcinoma and angiosarcoma. However, the role of microRNAs (miRNAs) in liver cell cycle changes under VC exposure remains unclear, which prevents research on the mechanism of VC-induced carcinogenesis. In this study, male rats were injected intraperitoneally with VC (0, 5, 25, and 125 mg/kg body weight) for 6, 8, and 12 weeks. Cell cycle analysis of liver cells, miRNA-222, miRNA-199a, miRNA-195, and miRNA-125b expression in the liver and serum, and target protein expression were performed at different time points. The results showed a higher percentage of hepatocytes in the G1/G0 and S phases at the end of 6 and 12 weeks of VC exposure, respectively. MiRNA-222 expression decreased initially and then increased, whereas miRNA-199a, miRNA-195, and miRNA-125b expression increased initially and then decreased, which corresponded with changes in cell cycle distribution and related target proteins expression (p27, cyclinA, cyclinD1, and CDK6). The corresponding expression levels of miRNAs in serum did not change. Dynamic changes in miR-222, miR-199a, miR-195, and miR-125b induced by VC can lead to cell cycle deregulation by affecting cell cycle-related proteins, and these miRNAs can serve as early biomarkers for malignant transformation caused by VC.

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A critical review on human serum Paraoxonase-1 in the literature: truths and misconceptions

Turkish Journal of Biochemistry ◽

10.1515/tjb-2020-0186 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Michael Mackness ◽

Eser Yildirim Sozmen

Keyword(s):

Heart Disease ◽

Experimental Design ◽

Human Serum ◽

Critical Review ◽

Neurological Diseases ◽

Rapid Expansion ◽

Paraoxonase 1 ◽

Serum Paraoxonase ◽

Inflammatory Component ◽

Biomedical Fields

AbstractHuman serum paraoxonase 1 (PON1) appears to play an important role in the development of a large variety of diseases with an inflammatory component including heart disease, diabetes, rheumatic diseases, neurological diseases and cancer. As such PON1 research is rapidly expanding into new biomedical fields. Unfortunately, this rapid expansion has resulted in a number of problems due to poor experimental design and the spreading of misconceptions in the literature. This review seeks to describe the basic properties of PON1 and the problems and misconceptions that have arisen.

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The inhibitory effect of silencing CDCA3 on migration and proliferation in bladder urothelial carcinoma

Cancer Cell International ◽

10.1186/s12935-021-01969-x ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Dexin Shen ◽

Yayun Fang ◽

Fenfang Zhou ◽

Zhao Deng ◽

Kaiyu Qian ◽

...

Keyword(s):

Cell Cycle ◽

Urothelial Carcinoma ◽

Carcinoma Cells ◽

Expression Level ◽

Tumor Cell Growth ◽

Migration Ability ◽

Bladder Urothelial Carcinoma ◽

Related Proteins

Abstract Background CDCA3 is an important component of the E3 ligase complex with SKP1 and CUL1, which could regulate the progress of cell mitosis. CDCA3 has been widely identified as a proto-oncogene in multiple human cancers, however, its role in promoting human bladder urothelial carcinoma has not been fully elucidated. Methods Bioinformatic methods were used to analyze the expression level of CDCA3 in human bladder urothelial carcinoma tissues and the relationship between its expression level and key clinical characteristics. In vitro studies were performed to validate the specific functions of CDCA3 in regulating cell proliferation, cell migration and cell cycle process. Alterations of related proteins was investigated by western blot assays. In vivo studies were constructed to validate whether silencing CDCA3 could inhibit the proliferation rate in mice model. Results Bioinformatic analysis revealed that CDCA3 was significantly up-regulated in bladder urothelial carcinoma samples and was related to key clinical characteristics, such as tumor grade and metastasis. Moreover, patients who had higher expression level of CDCA3 tend to show a shorter life span. In vitro studies revealed that silencing CDCA3 could impair the migration ability of tumor cells via down-regulating EMT-related proteins such as MMP9 and Vimentin and inhibit tumor cell growth via arresting cells in the G1 cell cycle phase through regulating cell cycle related proteins like p21. In vivo study confirmed that silencing CDCA3 could inhibit the proliferation of bladder urothelial carcinoma cells. Conclusions CDCA3 is an important oncogene that could strengthen the migration ability of bladder urothelial carcinoma cells and accelerate tumor cell growth via regulating cell cycle progress and is a potential biomarker of bladder urothelial carcinoma.

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In vivo/in silico insight into the effect of titanium dioxide nanoparticle on serum paraoxonase 1 activity in rat

Journal of Biomolecular Structure and Dynamics ◽

10.1080/07391102.2020.1864662 ◽

2021 ◽

pp. 1-11

Author(s):

Hessameddin Mortazavi ◽

Hossein Omidi-Ardali ◽

Seyed Asadollah Amini ◽

Javad Saffari-Chaleshtori ◽

Keihan Ghatreh Samani

Keyword(s):

Titanium Dioxide ◽

In Silico ◽

Paraoxonase 1 ◽

Titanium Dioxide Nanoparticle ◽

Serum Paraoxonase ◽

Insight Into

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Structure-Based Virtual Screening and Biological Evaluation of Peptide Inhibitors for Polo-Box Domain

Molecules ◽

10.3390/molecules25010107 ◽

2019 ◽

Vol 25 (1) ◽

pp. 107 ◽

Cited By ~ 1

Author(s):

Fang Yan ◽

Guangmei Liu ◽

Tingting Chen ◽

Xiaochen Fu ◽

Miao-Miao Niu

Keyword(s):

Cell Cycle ◽

Virtual Screening ◽

Fold Increase ◽

Pharmacophore Modeling ◽

Biological Evaluation ◽

Peptide Inhibitors ◽

Regulatory Proteins ◽

Competitive Binding Assay ◽

M Phase ◽

Cell Cycle Regulatory Proteins

The polo-box domain of polo-like kinase 1 (PLK1-PBD) is proved to have crucial roles in cell proliferation. Designing PLK1-PBD inhibitors is challenging due to their poor cellular penetration. In this study, we applied a virtual screening workflow based on a combination of structure-based pharmacophore modeling with molecular docking screening techniques, so as to discover potent PLK1-PBD peptide inhibitors. The resulting 9 virtual screening peptides showed affinities for PLK1-PBD in a competitive binding assay. In particular, peptide 5 exhibited an approximately 100-fold increase in inhibitory activity (IC50 = 70 nM), as compared with the control poloboxtide. Moreover, cell cycle experiments indicated that peptide 5 effectively inhibited the expression of p-Cdc25C and cell cycle regulatory proteins by affecting the function of PLK1-PBD, thereby inducing mitotic arrest at the G2/M phase. Overall, peptide 5 can serve as a potent lead for further investigation as PLK1-PBD inhibitors.

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