Proteomics Study of Mesenchymal Stem Cell-Like Cells Isolated from Cerebrospinal Fluid of Patients with Meningioma

2019 ◽  
Vol 16 (4) ◽  
pp. 282-288
Author(s):  
Arash Saffarian ◽  
Amir Tarokh ◽  
Mohammad Reza Haghshenas ◽  
Mousa Taghipour ◽  
Nooshafarin Chenari ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Cancer Progression ◽  
Flow Cytometer ◽  
Differentially Expressed ◽  
Phosphoglycerate Mutase ◽  
Intracranial Meningioma ◽  
Type I ◽  
Differentially Expressed Proteins ◽  
Isolated Cells ◽  
Proteomics Study

Background:Cerebrospinal fluid (CSF) contains pro-growth factors that can affect proliferation, migration and differentiation of Mesenchymal Stem Cells (MSCs).Objective:This study aimed to isolate MSC like cells from CSF of patients with meningioma and psudotumorcerebri (PTC) and identify differentially expressed proteins in these cells.Methods:Five patients with newly diagnosed intracranial meningioma and five patients with PTC were recruited in this comparative proteomics study. MSCs were isolated from CSF and validated by mesenchyml and non-mesenchyml fluorochrome antibodies, and flow cytometer analysis. Two- Dimensional Gel Electrophoresis (2-DE) coupled with Mass Spectrometry (MS) was performed to identify differentially expressed proteins.Results:Microscopic views of the isolated cells as well as flow cytometer analysis were found to be compatible with MSC-like cells. Eight distinct protein spots were differentially and reproducibly expressed among the stained gels of two studied groups. The identified proteins were Phosphoglycerate Mutase 1 (PGAM1), LIM and SH3 domain protein (LASP1), peroxiredoxin-6 (PRDX-6), type I cytoskeletal 9 (KRT9), Superoxide Dismutase (SOD), endoplasmin, Stathmin 1 (STMN1), and glutathione S-transferase (GST).Conclusion:This study provides new insights into the plausible role of CSF derived MSCs in cancer progression, and reveals a promising therapeutic opportunity for targeting of MSC proteins in patients with meningioma.

Identification of RIZ1 Targets Involvedin Erythroid Differentiation of K562 Human Erythroleukemia Cells Using Surface-Enhanced Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (SELDI).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2031-2031
Author(s):  
Naoto Takahashi ◽  
Matthew Brainbridge ◽  
Stuart A. Scott ◽  
Ryo Ichinohasama ◽  
Stephen E. Sanche ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Calcium Binding ◽  
Transforming Growth Factor ◽  
Erythroid Differentiation ◽  
Protein Chip ◽  
Differentially Expressed ◽  
Type I ◽  
Differentially Expressed Proteins ◽  
Erythroleukemia Cells ◽  
Tgf Β1

Abstract RIZ1 (PRDM2) is a member of the nuclear protein methyltransferase superfamily involved in chromatin remodeling. RIZ1 functions as a tumor suppressor gene in a number of human cancers and is down regulated in some human acute myeloid leukemias. We previously found RIZ-1 to be silenced in K562 erythroleukemia cells by promoter hypermethylation. Furthermore, expression of RIZ1 in K562 promotes erythroid differentiation and also potentiates TGF-β1 mediated differentiation. To investigate similarities between genes altered by RIZ1 expression and the TGF-β1 pathway, we used SELDI to compare the protein profiles of K562 against K562 + RIZ1 and K562 + TGF-β1. Protein extracts for SELDI profiling were separated into six fractions according to their isoelectric points. Proteins from each fraction were then bound to two different protein chip surfaces (H50-hydrophoboic and CM10-cation exhange) and their mass/charge determined using SELDI. We analyzed four replicates from each sample and classified proteins as differentially expressed if their P-values were below 0.05. In total, we observed 104 differentially expressed proteins (60 upregulated and 44 down regulated) between K562 and K562 + RIZ1 and 176 proteins (96 upregulated and 80 down regulated) between K562 and K562 + TGF-β1. We used 2D-PAGE to identify differentially expressed proteins identified by SELDI analysis and located 48 proteins that were over expressed in K562 + RIZ1 and K562 + TGF-β1 relative to K562. To establish whether these proteins were the same proteins observed using SELDI, we determined if the proteins had the same pI and molecular weight and if the gel-eluted proteins bound to the same protein chip surface with the same mass/charge. 15 of 48 proteins passed the above criteria and we determined their identities using Trypsin-based peptide mapping strategies with molecular weight and pI restrictions. We identified two candidate proteins (14-3-3ε and S100/A13) that are similarly over expressed in K562 + RIZ1 and K562 + TGF-β1. These proteins have been shown to be associated with TGF-β1 signaling. Schistosomal 14-3-3ε interacts with SmRK1, a divergent type I transforming growth factor β1 receptor (TR-I) present on the surface of adult parasites and also binds to and activates human TR-I. S100/A13 belongs to a family of low molecular weight proteins characterized by the presence of two calcium-binding EF-hand motifs that includes S100C/A11, a member recently shown to play a key role in a PKCα mediated pathway essential for the growth inhibition of normal human keratinocytes by TGF-β1. In summary, we demonstrate the potential for using SELDI to identify novel proteins involved in regulating and connecting cellular growth and differentiation pathways.

scholarly journals Proteomic Studies on the Mechanism of Myostatin Regulating Cattle Skeletal Muscle Development

2021 ◽  
Vol 12 ◽  
Author(s):  
Hui Sheng ◽  
Yiwen Guo ◽  
Linlin Zhang ◽  
Junxing Zhang ◽  
Manning Miao ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Matrix ◽  
Focal Adhesion ◽  
Growth And Development ◽  
Muscle Growth ◽  
Differentially Expressed ◽  
Type I ◽  
Differentially Expressed Proteins ◽  
Mstn Gene ◽  
Bovine Skeletal Muscle

Myostatin (MSTN) is an important negative regulator of muscle growth and development. In this study, we performed comparatively the proteomics analyses of gluteus tissues from MSTN+/− Mongolian cattle (MG.MSTN+/−) and wild type Mongolian cattle (MG.WT) using a shotgun-based tandem mass tag (TMT) 6-plex labeling method to investigate the regulation mechanism of MSTN on the growth and development of bovine skeletal muscle. A total of 1,950 proteins were identified in MG.MSTN+/− and MG.WT. Compared with MG.WT cattle, a total of 320 differentially expressed proteins were identified in MG.MSTN cattle, including 245 up-regulated differentially expressed proteins and 75 down-regulated differentially expressed proteins. Bioinformatics analysis showed that knockdown of the MSTN gene increased the expression of extracellular matrix and ribosome-related proteins, induced activation of focal adhesion, PI3K-AKT, and Ribosomal pathways. The results of proteomic analysis were verified by muscle tissue Western blot test and in vitro MSTN gene knockdown test, and it was found that knockdown MSTN gene expression could promote the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells (BSMSCs). At the same time, Co-Immunoprecipitation (CO-IP) assay showed that MSTN gene interacted with extracellular matrix related protein type I collagen α 1 (COL1A1), and knocking down the expression of COL1A1 could inhibit the activity of adhesion, PI3K-AKT and ribosome pathway, thus inhibit BSMSCs proliferation. These results suggest that the MSTN gene regulates focal adhesion, PI3K-AKT, and Ribosomal pathway through the COL1A1 gene. In general, this study provides new insights into the regulatory mechanism of MSTN involved in muscle growth and development.

Elevated Expression of A-Raf and FA2H in Hepatocellular Carcinoma is Associated with Lipid Metabolism Dysregulation and Cancer Progression

2019 ◽  
Vol 19 (2) ◽  
pp. 236-247 ◽  
Author(s):  
Maryam Ranjpour ◽  
Saima Wajid ◽  
Swatantra K. Jain
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cancer Progression ◽  
Early Stage ◽  
Tumor Stage ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
2D Electrophoresis ◽  
Differentially Expressed Proteins ◽  
Elevated Expression ◽  
Male Wistar Rats

Background:Identification of events leading to hepatocellular carcinoma (HCC) progression is essential for understanding its pathophysiology. The aims of this study are to identify and characterize differentially expressed proteins in serum of HCC-bearing rats and the corresponding controls during cancer initiation, progression and tumorigenesis.Methods:Chemical carcinogens, N-Nitrosodiethylamine and 2-aminoacetylfluorine are administered to induce HCC to male Wistar rats. The 2D-Electrophoresis and PD-Quest analyses are performed to identify several differentially expressed proteins in serum of HCC-bearing animals. These proteins are further characterized by MALDI-TOF-MS/MS analyses. Using pathwaylinker a HCC-specific network is analyzed among the MALDITOF- MS/MS characterized proteins and their interactors.Results:Carcinogen administration caused inflammation leading to liver injury and HCC development. Liver inflammation was confirmed by increase in the levels of TNF-α and IL-6 in carcinogen treated rats. We report significant increase in expression of two differentially expressed proteins, namely, A-Raf and Fatty Acid 2- Hydroxylase (FA2H), at early stage of HCC initiation, during its progression and at tumor stage. Real-time PCR analysis of mRNA for these proteins confirmed up-regulation of their transcripts. Further, we validated our experimental data with sera of clinically confirmed liver cancer patients.Conclusion:The study suggests that FA2H and A-Raf play a major role in the progression of HCC.

scholarly journals Biochemical Studies on Cerebrospinal Fluid in Patients With Unresponsive Wakefulness Syndrome: Toward a Potential Search for Biomarkers

2021 ◽  
Author(s):  
Lijuan Cheng ◽  
huang yuehong ◽  
Yan Luo ◽  
Hao Yang ◽  
Xiaohua Hu ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Dna Replication ◽  
Minimally Conscious State ◽  
Conscious State ◽  
Differentially Expressed ◽  
Control Group ◽  
Differentially Expressed Proteins ◽  
Unresponsive Wakefulness Syndrome ◽  
Minimally Conscious ◽  
Potential Biomarkers

Abstract Background: This study aimed to examine and screen patients for potential biomarkers for the diagnosis of unresponsive wakefulness syndrome (UWS). Methods: Patients with UWS, patients who regained consciousness (RC; patients in a minimally conscious state), and patients who had emerged from the minimally conscious state were evaluated using the Coma Recovery Scale-Revised (CRS-R). Cerebrospinal fluid (CSF) was collected from five healthy controls and 10 patients (5 UWS; 5 RC). Two-dimensional electrophoresis and proteomic analysis were used to examine and identify differentially expressed proteins in the CSF. Results: Compared with the control group, there were at least six proteins expressed at higher levels and two proteins expressed at lower levels in the CSF of the RC group than in the CSF of the UWS group. However, expression of vitamin D-binding protein (VDP) showed a further increase of 4.52-fold, that of DNA replication licensing factor mini-chromosome maintenance 4 (MCM4) showed a reduction of -20.61-fold, and that of hemoglobin subunit beta reversed to -8.34-fold in the UWS group. Another converse expression was of protein coenzyme Q-binding protein COQ10 homolog A (COQ10A), which was -4.31-fold in the RC group and 1.51-fold in the UWS group, compared to the control group. Expression of other proteins demonstrated an ascending trend in the RC group compared to the control group. Conclusions: The three differentially expressed proteins, VDP, DNA replication licensing factor MCM4, and hemoglobin subunit beta, may be involved in consciousness maintenance. This study identified potential biomarkers for UWS and will provide new insights into the pathogenesis of UWS.

scholarly journals Interleukin-1 in cerebrospinal fluid for evaluating the neurological outcome in traumatic brain injury

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Yingming Yue ◽  
Chongzhi Shang ◽  
Huajiang Dong ◽  
Kun Meng
Keyword(s):  
Traumatic Brain Injury ◽  
Cerebrospinal Fluid ◽  
Brain Injury ◽  
Neurological Outcome ◽  
Interleukin 1 ◽  
Differentially Expressed ◽  
Strongly Correlated ◽  
Differentially Expressed Proteins ◽  
Csf Proteins ◽  
Gcs Score

Abstract Objective Severe traumatic brain injury (TBI) is associated with unfavorable outcomes secondary to injury from activation of the inflammatory cascade, the release of excitotoxic neurotransmitters, and changes in the reactivity of cerebral vessels, causing ischemia. Inflammation induced by TBI is complex, individual-specific, and associated with morbidity and mortality. The aim of the present study was to discover the differentially expressed cerebrospinal fluid (CSF) proteins and identify which can improve the clinical outcomes in TBI patients. Methods In the present study, we reported 145 patients with TBI and found the change in patients’ leukocytes in serum and interleukin-1 (IL-1) in CSF, which strongly correlated with the neurological outcome. In terms of results of leukocytes in blood and IL-1 in CSF, we retained the patient’s CSF specimens and conducted a proteomic analysis. Results A total of 119 differentially expressed proteins were detected between samples of TBI and the normal, which were commonly expressed in all samples, indicating the differentially expressed proteins. When the patients’ Glasgow outcome score (GOS) improved, IL-1 was down-regulated, and when the patients’ GCS score deteriorated, IL-1 was up-regulated accompanied with the progression in TBI. Conclusion The differentially expressed proteins in CSF may be the novel therapeutic targets for TBI treatment. The leukocytes in blood samples and the IL-1 in CSF may be two important indicators for predicting the prognosis of TBI patients.

Mass Spectrometry-based Label-free Quantitative Proteomic Analysis of CCl4-induced Acute Liver Injury in Mice Intervened by Total Glycosides from Ligustri Lucidi Fructus

2020 ◽  
Vol 17 ◽  
Author(s):  
Qian Lu ◽  
Hai-Zhu Xing ◽  
Nian-Yun Yang
Keyword(s):  
Insulin Resistance ◽  
Liver Injury ◽  
Protein Expression ◽  
Signaling Pathway ◽  
Proteomic Analysis ◽  
Acute Liver Injury ◽  
Liver Cells ◽  
Differentially Expressed ◽  
Liver Protein ◽  
Differentially Expressed Proteins

Background: CCl4 acute liver injury (ALI) is a classical model for experimental research. However, there are few reports involved in the fundamental research of CCl4-induced ALI Ligustri Lucidi Fructus (LLF) are and its prescription have been used to treat hepatitis illness clinically. LLF and its active ingredients displayed anti-hepatitis effects, but the mechanism of function has not been fully clarified Objective: To investigate the proteomic analysis of CCl4-induced ALI, and examine the effects of active total glycosides (TG) from LLF on ALI of mice4, including histopathological survey and proteomic changes of liver tissues, and delineate the possible underlying mechanism. Methods: CCl4 was used to produce ALI mice model. The model mice were intragastrically administrated with TG and the liver his-topathological changes of mice were examined. At the end of test, mice liver samples were collected, after protein denaturation, re-duction, desalination and enzymatic hydrolysis, identification was carried out by nano LC-ESI-OrbiTrap MS/MS technology. The data was processed by Maxquant software. The differentially-expressed proteins were screened and identified, and their biological information was also analyzed based on GO and KEGG analysis. Key protein expression was validated by Western blot analysis Results: A total of 705 differentially-expressed proteins were identified during the normal, model and administration group. 9 signifi-cant differential proteins were focused based on analysis. Liver protein expression changes of CCl4-induced ALI mice were mainly involved in several important signal channels, namely FoxO signaling pathway, autophagy-animal, insulin signaling pathway. TG has anti-liver damnification effect in ALI mice, the mechanism of which is related to FoxO1 and autophagy pathways Conclusion: CCl4 inhibited expression of insulin-Like growth factor 1 (Igf1) and 3-phosphoinositide-dependent protein kinase 1 (Pdpk1) in liver cells and induced insulin resistance, thus interfered with mitochondrial autophagy and regeneration of liver cells and the metabolism of glucose and lipid, and caused hepatic necrosis in mice. TG resisted liver injury in mice. TG adjusted the expression level of key proteins Igf1 and Pdpk1 after liver injury and improved insulin resistance, thus promoted autophagy and resisted the liver damage

scholarly journals Differentially expressed proteins in platelets derived from patients with hypertension

2021 ◽  
Author(s):  
Yobana Armenta-Medina ◽  
Ivette Martínez-Vieyra ◽  
Oscar Medina-Contreras ◽  
Claudia G. Benitez-Cardoza ◽  
Albertana Jiménez-Pineda ◽  
...  
Keyword(s):  
Differentially Expressed ◽  
Differentially Expressed Proteins
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