(B) MiR-127-3p expression in 6 normal brain tissues (N, test was used to examine differences in miR-127-3p levels between normal brain tissues and GBM tissues (animal studies

(B) MiR-127-3p expression in 6 normal brain tissues (N, test was used to examine differences in miR-127-3p levels between normal brain tissues and GBM tissues (animal studies. GBM cell growth by inducing G1-phase arrest both and and value 0.05 was considered to (Glp1)-Apelin-13 be statistically significant. Results MiR-127-3p is significantly downregulated in GBM and is highly induced by DNA demethylation and histone deacetylase inhibition. Using next generation sequencing, we analyzed the miRNA profiles of four GBM and four normal brain tissues, and we found that miR-127-3p was significantly downregulated (reads per million from the next generation sequencing data. (B) MiR-127-3p expression in 6 normal brain tissues (N, test was used to examine differences in miR-127-3p levels between normal brain tissues and GBM tissues (animal studies. We found that miR-127-3p overexpression inhibited colony formation in both LN229 and T98 cells (growth of human GBM cells in nude mice. Open in a separate window FIG. 4. MiR-127-3p decreases GBM cell colony formationand inhibits GBM cell growth growth of human GBM cells in nude mice (Fig. 4). These data appear to suggest that miR-127-3p functions as a tumor suppressor. Our data are consistent with a recent publication by Guo et al. (2013) showing that ectopic expression of miR-127 in the gastric cancer cell line HGC-27 inhibits cell proliferation, cell cycle progression, and cell migration and invasion. Although Saito et al. (2006), Guo et al. (2013), and our group have shown that miR-127 expression is downregulated in GBM as well as gastric, prostate, bladder, and colon cancers, other groups have observed that miR-127-3p expression was higher in acute myeloid leukemia (Dixon-McIver et al., 2008), esophageal squamous cell carcinomas (Zhang et al., 2010), and cervical cancers (Lee et al., 2008). These data provide support for the hypothesis that the function of miR-127 is cancer type-specific. We also demonstrated for the first time that miR-127-3p targets five genes: SKI, RGMA, ZWINT, SERPINB9, and SFRP1 (Fig. 5), thus expanding the list of validated target genes of miR-127. The proto-oncogenes BCL6 (B-cell CLL/lymphoma 6) and MAPK4 (mitogen-activated protein kinase 4) were previously shown to be targets of miR-127 (Guo et al., 2013; Saito et al., 2006). Using the TCGA data, we analyzed the miR-127 expression with the survival of GBM patients, but did not find a significant association between the two. We also performed a survival analysis using the Rembrandt databases for the five target genes of miR-127: SKI, RGMA, ZWINT, SERPINB9, and SFRP1. We found only one gene whose expression is significantly associated with patient survival, considering only and em in vivo /em . Furthermore, we showed that SKI, RGMA, ZWINT, SERPINB9, and SFRP1 are the direct target genes of miR-127-3p. Finally, we found that miR-127-3p activates TGF- signaling to suppress GBM cell growth by targeting SKI in GBM. Supplementary Material Supplemental data:Click here to view.(22K, pdf) Supplemental data:Click here to view.(22K, pdf) Supplemental data:Click here to view.(18K, pdf) Supplemental data:Click here to view.(21K, pdf) Supplemental data:Click here to view.(69K, pdf) Supplemental data:Click here to view.(130K, pdf) Acknowledgments The work was funded by Grant 81072060 from the National Natural Science Foundation of China; Grants or loans PTGS2 2008DFA11320 and 2012AA022705 in the Ministry of Technology and (Glp1)-Apelin-13 Research, China; Offer 20110101120153 in the Ministry of Education, China; Offer 2012R10021 in the Zhejiang Provincial Federal government, and Offer 2011A11013 in the Shaoxing Main Technological and Scientific Tasks. The financing resources acquired no function within the scholarly research design and style; the collection, (Glp1)-Apelin-13 interpretation or evaluation of the info; the preparation from the manuscript; or your choice to send the manuscript for publication. Writer Disclosure Declaration The authors declare that no contending financial (Glp1)-Apelin-13 interests can be found..