scholarly journals Innovative finding of 266-nm laser regulating CD90 levels in SDSCs

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ray-Ling Hsiao ◽  
Yen-Chung Chen ◽  
Mei-Yue Huang ◽  
Chiang-Yun Chen ◽  
Yu-Wei Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Light Intensity ◽  
Protein Expression ◽  
Cellular Protein ◽  
Maximal Effect ◽  
Cell Markers ◽  
Regulate Protein ◽  
Protein Alterations ◽  
Irradiate Skin

AbstractWe used light to irradiate skin-derived stem cells and tried to find any cellular protein alterations 24 h after illumination. A 266-nm laser with four intensities was used, and of the nine cell markers that were surveyed in our trials, only CD90 was downregulated at an intensity of 20 μJ for 10 s. Repeated illuminations from the 266-nm laser at seven intensities revealed that CD90 expression was downregulated 14.6–28.8%, depending on light intensity. The maximal effect was noted at an intensity of 30 μJ for 2 s. This innovative finding reveals that a 266-nm laser can regulate protein expression in skin-derivative stem cells.

scholarly journals TMOD-18. TARGETING THE PI3K/AKT PATHWAY IN MYCN AMPLIFIED HIGH GRADE GLIOMAS

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii231-ii232
Author(s):  
Katharine Halligan ◽  
Ann-Catherine Stanton ◽  
Matthew Halbert ◽  
Brian Golbourn ◽  
Stephen Mack ◽  
...  
Keyword(s):  
Stem Cells ◽  
Tumor Suppressor ◽  
Mouse Model ◽  
Protein Expression ◽  
Neural Stem Cells ◽  
Tumor Suppressor Genes ◽  
Akt Pathway ◽  
High Grade ◽  
Suppressor Genes ◽  
High Grade Gliomas

Abstract Pediatric glioblastoma (pGBM) are incurable brain tumors with overall poor prognosis and response to treatments due to molecular and epigenetic heterogeneity. In particular, the MYCN subtype of pGBM are a highly aggressive form of GBM with a dismal median survival of only 14 months. Furthermore, this subtype is enriched with loss of the tumor suppressor genes TP53 and PTEN, leading to aberrantly active PI3K-AKT signaling pathway and DNA-checkpoint abnormalities. Here, we report the generation of a novel syngeneic mouse model that recapitulates the features of the MYCN subtype of pGBM. We isolated Sox2-Cre neural stem cells from C57BL/6 mice and transduced inverted retroviral-cassettes of the murine Mycn oncogene simultaneously with shRNA targeting tumor suppressor genes p53 and Pten. Retroviral-cassettes are flanked by tandem LoxP sites arranged so that Cre recombinase expression inverts the cassettes in frame allowing for MYCN protein expression and loss of the P53/PTEN proteins. Transgene activation is accompanied with selectable cell surface markers and fluorescent tags enabling for fluorescent activated cell sorting (FACS) of the desired cell populations. Neural stem cells with MYCN protein expression and concurrent silencing of P53 and PTEN protein (NPP cells) result in significantly increased proliferation and activation of PI3K-AKT pathway as compared to control neural stem cells and have. Injection of NPP cells into the forebrain of immune competent C57BL/6 mice result in the formation of invasive high-grade gliomas with a lethal phenotype at ~50 days post injection. Using several next generation brain penetrant small molecule inhibitors of the PI3K-AKT pathway, we show inhibition of tumorigenesis in vitro. Moreover, we have identified several novel mechanisms of PI3KAKT treatment resistance and are currently identifying therapies that may overcome this resistance through RNA seq analysis. In summary, well defined genetic drivers of GBM can lead to informed mouse model generation to test promising therapies.

scholarly journals Translational repression of SLC26A3 by miR-494 in intestinal epithelial cells

2014 ◽  
Vol 306 (2) ◽  
pp. G123-G131 ◽  
Author(s):  
Arivarasu N. Anbazhagan ◽  
Shubha Priyamvada ◽  
Anoop Kumar ◽  
Daniel B. Maher ◽  
Alip Borthakur ◽  
...  
Keyword(s):  
Protein Expression ◽  
Therapeutic Strategy ◽  
Luciferase Activity ◽  
Negative Control ◽  
Intestinal Epithelial ◽  
Diarrheal Diseases ◽  
Regulate Protein ◽  
Putative Binding Site ◽  
Mammalian Intestine ◽  
Relative Luciferase Activity

SLC26A3 [downregulated in adenoma (DRA)] is a Cl−/HCO3− exchanger involved in electroneutral NaCl absorption in the mammalian intestine. Altered DRA expression levels are associated with infectious and inflammatory diarrheal diseases. Therefore, it is critical to understand the regulation of DRA expression. MicroRNAs (miRNAs) are endogenous, small RNAs that regulate protein expression via blocking the translation and/or promoting mRNA degradation. To investigate potential modulation of DRA expression by miRNA, five different in silico algorithms were used to predict the miRNAs that target DRA. Of these miRNAs, miR-494 was shown to have a highly conserved putative binding site in the DRA 3′-untranslated region (3′-UTR) compared with other DRA-targeting miRNAs in vertebrates. Transfection with pmirGLO dual luciferase vector containing DRA 3′-UTR (pmirGLO-3′-UTR DRA) resulted in a significant decrease in relative luciferase activity compared with empty vector. Cotransfection of the DRA 3′-UTR luciferase vector with a miR-494 mimic further decreased luciferase activity compared with cells transfected with negative control. The transfection of a miR-494 mimic into Caco-2 and T-84 cells significantly increased the expression of miR-494 and concomitantly decreased the DRA protein expression. Mutation of the seed sequences for miR-494 in 3′-UTR of DRA abrogated the effect of miR-494 on 3′-UTR. These data demonstrate a novel regulatory mechanism of DRA expression via miR-494 and indicate that targeting this microRNA may serve to be a potential therapeutic strategy for diarrheal diseases.

Characteristics of Porcine Induced Pluripotent Stem Cells in their MHC Protein Expression

2012 ◽  
Vol 94 (10S) ◽  
pp. 605
Author(s):  
K.-M. Park ◽  
H.-S. Nam ◽  
S.-M. Park ◽  
S.-H. Cha ◽  
I.-H. Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Protein Expression ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Induced Pluripotent

scholarly journals Protein Expression of Mesenchymal Stem Cells after Transfection of pcDNA3.1−-hVEGF165by Ultrasound-Targeted Microbubble Destruction

2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Zhaoxia Pu ◽  
Xiangdong You ◽  
Qiyuan Xu ◽  
Feng Gao ◽  
Xiaojie Xie ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Specific Gene ◽  
Elisa Kit ◽  
The Third ◽  
Marrow Mesenchymal Stem Cells ◽  
Organ Specific ◽  
A New Technique

Ultrasound-targeted microbubble destruction (UTMD) has been proposed as a new technique for organ-specific gene transfer and drug delivery. This study was performed to investigate the effect of UTMD on marrow mesenchymal stem cells (MSCs) transfected with pcDNA3.1−-hVEGF165.pcDNA3.1−-hVEGF165were transfected into the third passage of MSCs, with or without UTMD under different ultrasound conditions. Protein expression was quantified by hVEGF165-ELISA kit after transfection for 24, 48, and 72 hours. UTMD-mediated transfection of MSCs yielded a significant protein expression. UTMD of mechanic index (MI) 0.6 for 90 seconds led to the highest level of protein expression.

scholarly journals Prostate stem cells: The niche and cell markers

2008 ◽  
Vol 15 (4) ◽  
pp. 289-294 ◽  
Author(s):  
Tetsuya Takao ◽  
Akira Tsujimura
Keyword(s):  
Stem Cells ◽  
Cell Markers

Stress resistant human embryonic stem cells as a potential source for the identification of novel cancer stem cell markers

2010 ◽  
Vol 289 (2) ◽  
pp. 208-216 ◽  
Author(s):  
Shaker A. Mousa ◽  
Thangirala Sudha ◽  
Evgeny Dyskin ◽  
Usawadee Dier ◽  
Christine Gallati ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Cancer Stem Cell ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Stem Cell Markers ◽  
Cell Markers ◽  
Cancer Stem Cell Markers ◽  
Potential Source

Abstract 294: Expression of Matrix Metalloproteases during the Differentiation of Porcine Adopse-Drived Mesenchymal Stem Cells ro Endothelial Cells

2013 ◽  
Vol 33 (suppl_1) ◽  
Author(s):  
Sami G Almalki ◽  
Velidi Rao ◽  
Divya Pankajakshan ◽  
Devendra K Agrawal
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Mesenchymal Stem Cells ◽  
Enzyme Activity ◽  
Protein Expression ◽  
Western Blot ◽  
Gelatin Zymography ◽  
Matrix Metalloproteases ◽  
Positive Staining ◽  
Mrna Transcripts

Rationale Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent cells that have the potential to differentiate into different cell linages, and represent promising tools in various clinical applications. However, the molecular mechanisms that control the ability of ADMSCs to remodel 3-dimensional extracellular matrix (ECM) barriers during differentiation are not clearly understood. Herein, we studied the expression of matrix metalloproteinases (MMPs) during the differentiation of ADMSCs to endothelial cells (ECs) in vitro . Methods MSCs were isolated from porcine abdominal adipose tissue, and characterized by positive staining for MSC markers, CD44, CD73, CD90, and negative staining for CD11b, CD34 and CD45. The plasticity of MSCs was detected by bi-lineage differentiation to osteocytes, and adipocytes. The mRNA transcripts for different MMPs and TIMPs and protein expression of EC markers were analyzed by RT-PCR and immunostaining. The enzyme activity and protein expression were also analyzed by gelatin zymography, ELISA, and Western blot. Results The differentiation of ADMSCs to ECs was confirmed by the positive staining and mRNA expression of the endothelial markers. The mRNA transcripts for MMP-2 and membrane type 1 MMP (MT1-MMP) was significantly increased by 2.5 and 2.0 fold, respectively, during the differentiation of MSCs into ECs. Western blot and ELISA showed an elevated MT1-MMP and MMP-2 expression. The enzyme activity of MMP-2 was also observed by gelatin zymography. Conclusion We demonstrated that porcine ADMSCs have the ability to differentiate into ECs, and this process involves the up-regulation of MMP-2 and MT1-MMP. The increase in the expression of MMP-2 and MT1-MMP may, at least partially, facilitate the change in morphology of MSCs by degrading the ECM barriers. These findings may provide a potential mechanism for the role of MMP2 and MT1-MMP in the differentiation of ADMSCs into ECs.

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