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Mark Komarov
Mark Komarov

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Differentially expressed genes were also analyzed for significant enrichment with respect to functional categories using DAVID. The top KEGG pathways identified were cytokine-cytokine receptor interaction, chemokine signaling pathway, extracellular matrix (ECM)-receptor interaction, cell adhesion and focal adhesion. Enrichment of these functional categories was prominent in the CP-NCCIT vs. NP-NCCIT datasets. Of particular interest were the genes that contribute to the functioning and maintenance of ECM. Matrix metalloproteinases (MMPs) are involved in the degradation of ECM proteins and have been associated with tumor cell invasion. The membrane-anchored reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a potent inhibitor of MMP activity. RECK down-regulation has been identified in many cancers, and it has been reported that high RECK expression levels were associated with favorable prognosis in prostate cancer [26, 27]. In comparing the CP-NCCIT and NP-NCCIT expression profiles, several genes associated with RECK were differentially expressed (Fig. 2a). For example, induction of MMP9, a potential prostate cancer urine biomarker [28], was greater in CP-NCCIT than in NP-NCCIT (Fig. 2b). MMP9 expression was also higher in sorted CP vs. NP stromal cell transcriptomes. In contrast, RECK was more up-regulated in NP-NCCIT than CP-NCCIT, as was tissue inhibitor of metalloproteinase TIMP1, an antagonist of MMPs. With regard to the possible genesis of CP stromal cells, we also examined the effect of NCCIT factors on NP stromal cells [15]. NCCIT extracts, when injected into differentiated cells, can activate expression of stem cell genes [29]. Instead of extract injection, NCCIT factors were examined in the co-culture format with NP stromal cells. We found that at day 3, the co-cultured NP stromal cells showed a gene expression profile for both mRNA and microRNA resembling that of CP stromal cells. Thus, CP stromal cells appear to represent a more primitive cell type in the stromal lineage. This is certainly in line with their lower expression of smooth muscle cell genes, and CP stromal cells are characterized by a loss of smooth muscle differentiation [3]. The basal epithelium also contains the progenitor cell population, which could affect stromal cell differentiation. To isolate enough CD90+ NP stromal cells for study presents a technical challenge because of their low number, which necessitates the need to obtain large tissue specimens for sorting.




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With regard to the possible genesis of CP stromal cells, we also examined the effect of NCCIT factors on NP stromal cells [15]. NCCIT extracts, when injected into differentiated cells, can activate expression of stem cell genes [29]. Instead of extract injection, NCCIT factors were examined in the co-culture format with NP stromal cells. We found that at day 3, the co-cultured NP stromal cells showed a gene expression profile for both mRNA and microRNA resembling that of CP stromal cells. Thus, CP stromal cells appear to represent a more primitive cell type in the stromal lineage. This is certainly in line with their lower expression of smooth muscle cell genes, and CP stromal cells are characterized by a loss of smooth muscle differentiation [3]. The basal epithelium also contains the progenitor cell population, which could affect stromal cell differentiation. To isolate enough CD90+ NP stromal cells for study presents a technical challenge because of their low number, which necessitates the need to obtain large tissue specimens for sorting.


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