Supplementary MaterialsS1 Fig: The correlation between miR-513a and NEDD4L expression levels in TCGA microarray data

Supplementary MaterialsS1 Fig: The correlation between miR-513a and NEDD4L expression levels in TCGA microarray data. in U87 MG cells (log2 (Ratio) Q-0.58; adj. value Q0.05). (PDF) pone.0225913.s005.pdf (304K) GUID:?E28139ED-AEC6-466D-8B58-80E0CBBC1B95 S1 File: Normalized microarray data. (XLSX) pone.0225913.s006.xlsx (132K) GUID:?949B061F-9393-4D85-A4BB-F87B1221854A Attachment: Submitted filename: gene locus was upregulated by IGF-1-mediated phosphoinositide 3-kinase (PI3K) signaling. Its elevated levels were also observed in gliomas versus normal cells, in array data of The Malignancy Genome Atlas (TCGA), and the “type”:”entrez-geo”,”attrs”:”text”:”GSE61710″,”term_id”:”61710″GSE61710, “type”:”entrez-geo”,”attrs”:”text”:”GSE37366″,”term_id”:”37366″GSE37366, and “type”:”entrez-geo”,”attrs”:”text”:”GSE41032″,”term_id”:”41032″GSE41032 datasets. In addition, lower levels of neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L), an E3 ubiquitin protein ligase that inhibits WNT signaling, were found in gliomas by analyzing cells, arrays, and RNA sequencing data of TCGA glioma patients. Furthermore, a negative correlation was identified between miR-513a-5p and NEDD4L in glioma. NEDD4L was also validated as a direct target gene of miR-513a-5p, and it was reduced by IGF-1 treatment. Overexpression of NEDD4L inhibited glioma cell viability and reversed IGF-1-repressed TMZ cytotoxicity. In contrast, miR-513a-5p Chaetominine significantly affected NEDD4L-inhibited WNT signaling and reduced TMZ cytotoxicity. These findings demonstrate a distinct role of IGF-1 signaling through miR-513a-5p-inhibited NEDD4L networks in influencing GBM’s drug sensitivity to TMZ. Introduction Glioblastoma Chaetominine multiforme (GBM) belongs to grade IV primary malignant gliomas with poor prognoses and high lethality in adults [1, 2]. Several tumor microenvironmental factors were identified to enhance the risk of brain tumors, including the insulin-like growth factor (IGF) signaling axis [3]. When the circulating IGF-1 ligand binds to its receptor, IGF-1R, this tyrosine kinase receptor is usually activated through an autophosphorylation mechanism. Subsequently, two major downstream pathways, phosphoinositide 3-kinase (PI3K)/AKT and Ras/extracellular signal-regulated kinase (ERK) pathways, are enhanced to prevent cell death or promote cell growth. In gliomas, IGF-1 modulates cell proliferation and strongly stimulates cell Chaetominine migration [4]. IGF-1 also regulates inflammatory responses in glioma cells via influencing hypoxia-inducible factor (HIF)-1-toll-like receptor 9 (TLR9) cross talk Chaetominine [5]. Furthermore, increasing evidence suggests that IGF-1 signaling is usually involved in drug resistance mechanisms, resulting in glioma progression [6]. The IGF-1/IGF-1R axis was identified to underlie resistance to colony-stimulating factor-1 receptor (CSF-1R) inhibition in gliomas [7]. By increasing Bcl-2 expression and decreasing caspase-3 protease activity, IGF-1 significantly decreased the etoposide-induced apoptosis of glioma cells [8]. Taken together, comprehensively investigating IGF-1-mediated gene networks may be helpful in understanding the development of gliomagenesis and offer innovative therapeutic approaches for glioblastomas. Micro (mi)RNAs are endogenous, little, non-coding RNAs that inhibit gene expressions by binding towards the 3 untranslated area (UTR) of their focus on messenger (m)RNAs. Aberrant miRNA expressions had been determined in GBM advancement [9]. For instance, miR-10b, a portrayed onco-miR in every GBM subtypes extremely, was suggested to be a potential focus on for GBM therapy [10]. Elevation of miR-215 amounts by hypoxia is essential for reprogramming glioma-initiating cells in GBM incident and recurrence [11]. miR-513a-5p, an intergenic miRNA, comes from two different gene loci: miR-513a-1 and miR-513a-2. The functions of miR-513a in tumorigenesis are still unclear, especially in GBM. Only one study reported that upregulated miR-513a-5p levels were observed in MGC18216 GBM patients compared to controls [12]. The functions and molecular mechanisms of miR-513a-5p in glioma progression need to be further analyzed. Neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L, also known as NEDD4-2) is an E3 ubiquitin protein ligase belonging to the NEDD4 family and contains a homologous E6-associated protein C-terminus (HECT) domain name [13]. The best known function of NEDD4L is as an ion channel regulator, including the epithelial sodium channel (ENaC) [14], Na+-Cl? cotransporter (NCC) [15], voltage-gated sodium channels (Navs) [16], and so on. Recently, a role of NEDD4L in carcinogenesis was recognized. NEDD4L negatively regulates canonical WNT signaling in colorectal malignancy [17]. Decreased NEDD4L levels were correlated with poor prognoses in gastric malignancy patients [18]. Similarly, in gliomas, reduced NEDD4L expression Chaetominine was associated with aggressive progression and worse prognoses [19], suggesting that NEDD4L could be a tumor suppressor that inhibits tumorigenesis. However, no other studies.