Introduction Chordoma is a rare, resistant bone tumour thought to be arised from remnants of embryonic notochord. Cancer stem cells (CSCs) are associated with tumorigenesis, recurrence and resistance in cancers. Therefore, chordoma CSCs are possible targets for chordoma treatment. According to our previous study, CD133 and CD15 expressing chordoma cells shows stem-like cell properties however miRNA and mRNA profiling of these cells have not been determined yet. Material and methods In this study, we determined dysregulated miRNAs in CSCs of chordoma via miRNA microarray and validated these miRNAs with qPCR. Hereupon, validated miRNAs was transfected chordoma cell lines transiently and identified their role in proliferation, apoptosis, migration and invasion capacities. mRNA microarray analysis was also carried out to examine dysregulated pathways in chordoma CSCs. Lastly, relationship between clinicopathological features and dysregulated miRNAs (and their targets) has been evaluated in 21 chordoma patients Results and discussions CD133 +CD15+cells in chordoma cell lines exhibits CSC phenotype with increased CSC-related gene expression and invasion-migration ability. miRNA microarray analyses indicated that 238 miRNAs (15 upregulated, 223 downregulated) were differentially expressed in chordoma Cancer Stem Cells (CSCs). mRNA microarray results also suggested that chordoma CSCs differentially expressed 1064 genes (176 upregulated, 888 downregulated). We then demonstrated that Wnt5a, TGF-α, Btg2 and Mycbp genes which are relevant to CSC-related pathways were direct targets of miR-140–3 p, miR-148a-3p, miR-210–5 p and miR-574–5 p, respectively. Finally, we determined that miR-140–3 p and miR-148a-3p expression was correlated with Ki67, miR-140–3 p and TGFa expressions was correlated with p53, and also MYCBP expression was positively correlated with tumour volume. Finally, mRNA microarray analysis showed that various pathways including Cell cycle, DNA replication, Spliceosome, Mismatch repair, FoxO signalling, P53 Signalling, Hippo signalling, BRCA2 and ATR in Cancer Susceptibility, and Apoptotic DNA fragmentation and tissue homeostasis were dysregulated in chordoma CSCs. Conclusion Through these findings, we concluded that chordoma CSCs have distinctive miRNA and mRNA profiles and this situation can regulate stemness properties of chordoma CSCs.
Hematopoietic stem cells (HSCs) are particularly characterized by their quiescence and self-renewal. Cell cycle regulators tightly control quiescence and self-renewal capacity. Studies suggest that modulation of ubiquitination and neddylation could contribute to HSC function via cyclin-dependent kinase inhibitors (CDKIs). S-phase kinase-associated protein 2 (SKP2) is responsible for ubiquitin-mediated proteolysis of CDKIs. Here, we modulated overall neddylation and SKP2-associated ubiquitination in HSCs by using SKP2-C25, an SKP2 inhibitor, and MLN4924 (Pevonedistat) as an inhibitor of the NEDD8 system. Treatments of SKP2-C25 and MLN4924 increased both murine and human stem and progenitor cell (HSPC) compartments. This is associated with the improved quiescence of murine HSC by upregulation of p27 and p57 CDKIs. A colony-forming unit assay showed an enhanced in vitro self-renewal potential post inhibition of ubiquitination and neddylation. In addition, MLN4924 triggered the mobilization of bone marrow HSPCs to peripheral blood. Intriguingly, MLN4924 treatment could decrease the proliferation of murine bone marrow mesenchymal stem cells or endothelial cells. These findings shed light on the contribution of SKP2, and associated ubiquitination and neddylation in HSC maintenance, self-renewal, and expansion.
<p>FH is expressed in human glioma and correlates with disease severity. FH is produced in human lower-grade glioma (<b>A</b>) and GBM (<b>B</b>). <b>C,</b> FH expression correlates with decreased overall survival (<b>C</b>) and disease-free survival (<b>D</b>) of patients with glioma. Expression of FH correlates with primary therapy outcome (<b>E</b>), cancer type (<b>F</b>), neoplasm histological grade (<b>G</b>), new neoplasm event post-initial therapy (<b>H</b>), and <i>IDH1</i> mutation rate (<b>I</b>). The worse survival rendered by FH is ICOS-dependent (<b>J</b>) but not dependent on ICOSL (<b>K</b>). The survival estimates and correlation with other clinical parameters based on mRNA and survival data of <i>n</i> = 509 (<b>C</b>, <b>J</b>, and <b>K</b>), <i>n</i> = 472 (<b>D</b>) patients from TCGA provisional dataset brain lower-grade glioma, analyzed with cBioPortal. Statistical tests: logrank test (<b>C</b>, <b>D</b>, <b>J</b>, and <b>K</b>), <i>χ</i><sup>2</sup> test (<b>E–I</b>). Representative picture of <i>n</i> = 2 (<b>A</b>), <i>n</i> = 3 patients (<b>B</b>). </p>
c-Myc plays a major role in the maintenance of glycolytic metabolism and hematopoietic stem cell (HSC) quiescence. Targeting modulators of HSC quiescence and metabolism could lead to HSC cell cycle entry with concomitant expansion. Here we show that c-Myc inhibitor 10074-G5 treatment leads to 2-fold increase in murine LSKCD34low HSC compartment post 7 days. In addition, c-Myc inhibition increases CD34+ and CD133+ human HSC number. c-Myc inhibition leads to downregulation of glycolytic and cyclin-dependent kinase inhibitor (CDKI) gene expression ex vivo and in vivo. In addition, c-Myc inhibition upregulates major HDR modulator Rad51 expression in hematopoietic cells. Besides, c-Myc inhibition does not alter proliferation kinetics of endothelial cells, fibroblasts or adipose derived mesenchymal stem cells, however; it limits bone marrow derived mesenchymal stem cell proliferation. We further demonstrate that a cocktail of c-Myc inhibitor 10074-G5 along with tauroursodeoxycholic acid (TUDCA) and i-NOS inhibitor L-NIL provides a robust HSC maintenance and expansion ex vivo as evident by induction of all stem cell antigens analyzed. Intriguingly, the cocktail of c-Myc inhibitor 10074-G5, TUDCA and L-NIL improves HDR related gene expression. These findings provide tools to improve ex vivo HSC maintenance and expansion, autologous HSC transplantation and gene editing through modulation of HSC glycolytic and HDR pathways.
Although meningioma is a common disease, there is a lack of understanding of the underlying molecular mechanisms behind its initiation and progression. We used combined miRNA-mRNA transcriptome analysis to discover dysregulated genes and networks in meningiomas. Fourteen fresh-frozen meningioma samples and one human meningeal cell line were analyzed by using miRNA and whole transcriptome microarray chips. Data was filtered and analyzed. Candidate miRNAs and mRNAs were selected for validation in fifty-eight patient samples. miRNA and target mRNA relationships were assessed by inhibiting miRNA in meningioma cells. Apoptosis and viability assays were also used as functional tests. With the whole transcriptome microarray, 3753 genes were found to be dysregulated, and 891 miRNAs were found to be dysregulated as a result of miRNA microarray. Results were combined and analyzed with bioinformatics tools. Top differential pathways included those of inflammation, cancer, and cellular growth and survival. The oncosupressor PTX3 was constitutively low in meningioma samples. Moreover, PTX3 negatively correlated with miR-29c in our samples. Inhibiting miR-29c upregulated the PTX3 level, induced apoptosis of meningioma cells, and decreased cell viability. CABIN1, miR-29c, TMOD1, PTX3, RPL22, SPARCL1 and RELA were correlated with clinicopathological features in patient samples. Our results present the first integrated mRNA-miRNA analysis in meningiomas. miR-29c-3p and PTX3 are inversely correlated in tissues and meningioma cells, hinting that PTX3 can be regulated by miR-29c-3p. Furthermore, we determined potential clinicopathological markers.
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