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Accordingly, we wanted to investigate whether an up-regulation of miR-362-3p in colon cancer cell lines would have a negative impact on http://www.selleck.cn/products/VX-809.html the viability of colon cancer cell lines. A recent report indicated that miR-362-3p was expressed by normal and cancerous colonocytes,[32] thereby justifying ectopic expression experiments in CRC cell lines. Subsequently, three different colon cancer cell lines (HCT116, LS174T and DLD1) were transfected with miR-362-3p. Transfection efficiency was evaluated using Cy3-labelled-scr miRNA control and fluorescence microscopy and was consistently above 80% in all cell lines (data not shown). Accordingly, RT-qPCR demonstrated significantly increased miR-362-3p levels 48 hours post transfection (Supporting Information Fig. S3). Ectopic expression of miR-362-3p resulted in significant reduced viability (cell viability reduction>25% and p http://www.selleckchem.com/products/lgk-974.html the finding by demonstrating repressed growth of miR-362-3p transfected cells (Fig. 2b). Furthermore, the presence of a miR-362-3p inhibitor could partially rescue cells from the miR-362-3p mediated reduction of cell viability (Fig. 2c), establishing a specific effect of the miRNA. A migration assay was carried out to assess further the functional role of elevated miR-362-3p in tumors from patients with no recurrence. A modest but significant inhibitory effect of miR-362-3p on the migration of DLD1 cells was identified adding further evidence to its role http://www.selleckchem.com/products/17-AAG(Geldanamycin).html as a tumor suppressor (Fig. 2d). The migration of non-treated DLD1 cells is shown in Supporting Information Figure S4A. In conclusion, ectopic miR-362-3p expression induces repression of cellular growth and has an inhibitory effect on cell migration. To elucidate whether the growth suppression of miR-362-3p was due to cellular death or cell cycle arrest, we performed Lactate Dehydrogenase (LDH) assays and cell cycle analyses. We observed no cellular death in response to ectopic expression of miR-362-3p (Fig. 2e). By contrast, a significant reduction of cells in the S phase (p=0.001) and an increased number of cells in the G1 phase (p=0.001) was induced (Fig. 2f). These results indicate that the reduced growth rate induced by miR-362-3p was at least partly caused by a G1/S arrest. To elucidate the mechanism behind the growth inhibitory effect of miR-362-3p we set out to identify miR-362-3p targets. Genome wide microarray transcription profiling of HCT116 cells over-expressing miR-362-3p was carried out to screen for target candidates regulated at the transcriptional level.