Error bars represent SDs (*, 0.05; **, 0.001). Next, siRNA targeting the autophagic key protein was used to further investigate whether the effects around the above phenotypes were achieved through autophagy. overexpression, knockdown promoted infiltration of U87 cells to the surrounding structures in nude mice in vivo. The above phenotypic changes were also observed in and HCT116 colon cancer cells. In summary, our study provided support for a link between autophagy and EMT status in WT GBM cells and provided evidence for the signaling pathway (exhibit significantly elevated expression in supratentorial primitive neuroectodermal tumor samples with amplification. In preliminary experiments, and have been shown to be upregulated in the adult brain as compared with that in neural stem cells (NSCs) and the fetal brain, suggesting that and may have important biological functions in the embryonic development of the brain (Fig. S1A). A literature review shows that, amplification is found in several other human tumors as well.20 Moreover, the cluster is a novel prognostic biomarker in hepatocellular carcinoma by miRNA profiling.21 In another publication, downregulation of and promotes the proliferation of hepatocellular carcinoma cells.22 In estrogen receptor (ER)-positive breast malignancy, about 50 of the miRNAs within the cluster were found to be significantly upregulated in chemoresistant cells. Among these miRNAs, regulates cell viability and cell cycle progression.23 Additionally, the expression of miRNAs is higher in villous trophoblasts than in extravillous trophoblasts, and both and regulate the migration of human trophoblasts.24 These reports support that and harbored in may play an important role in tumorigenesis in several types of human tumors. Since these 2 miRNAs may also have functions in the HMOX1 embryonic development of the brain, we aimed to determine whether the expression levels of these miRNAs were altered in 6H05 (trifluoroacetate salt) gliomas. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) analysis of 6 malignant glioma cell lines revealed that and were upregulated significantly in 2 lines (Fig. S1B), suggesting that these 2 miRNAs may also have functions in GBM. Moreover, as mentioned above, autophagy and the EMT are thought to be crucial in the progression of GBM. Therefore, in this study, we sought to comprehensively analyze the precise biological functions of these 2 miRNAs in the regulation of autophagy and the EMT phenotype in GBM cells in vitro and in vivo. Results Low expression was closely associated with poor prognosis in patients with GBM In order to validate the relevance of in the prognosis of patients with glioma, we examined the expression of and in patients with newly diagnosed GBM who had undergone surgery plus concomitant and adjuvant TMZ chemoradiotherapy (Stupp regimen: radiotherapy plus concomitant TMZ 75?mg/m2; adjuvant TMZ 150?mg/m2 5/28 d for 6 cycles)1 as recommended by the National Comprehensive Malignancy Network (NCCN) Clinical Practice Guideline for Central Nervous System Cancer (Version 2, 2014). Forty-six clinical samples were used for analysis of and by qRT-PCR, as shown in Physique 1A. The expression level of the 2 2 miRNAs together with the other variables, such as the patients age, gender, tumor volume and location (i.e., functional area or not), and degree of tumor resection were recorded (Table S1). Notably, in 23 cases (50%), was expressed at a level higher than the median; we defined this as the high-expression group or (+). The other 23 cases were included in the low-expression group or (-). Open in a separate window Physique 1. Low expression was associated with poor prognosis in patients with GBM. (A) and expression levels in 46 GBM samples were detected by qRT-PCR (using the 2-CT method); 23 cases (50%) exhibited high expression of these miRNAs, for which the expression was higher than the median (indicated by the red arrows; see also Tables S1 and S2). (B) Kaplan-Meier tumor-free survival analysis according to levels. Patients with low expression of (= 0.0014). (C) Kaplan-Meier overall survival analysis. The 23 patients with low expression had significantly poorer outcomes than those with high expression (= 0.0083). By 6H05 (trifluoroacetate salt) Kaplan-Meier analysis of each variable, tumor volume, functional area, expression level, and expression level were selected for Cox multivariate regression analysis of overall survival (OS). The functional area, degree of 6H05 (trifluoroacetate salt) tumor resection, and expression levels of and were selected for Cox regression analysis of progression-free survival (PFS). The final Cox multivariate regression analysis indicated that expression level affected the prognosis of OS (= 0.023) and PFS (= 0.003; Table S2). Moreover, PFS and OS curves consistently showed that patients in the (+) group exhibited better prognoses than patients in the (-) group (Fig. 1B and C). Induction of autophagy by TMZ was.