microarray Analysis of MKN-45 cell lines after 48 hours post-transfection of miR-125b mimic (50nM) and mimic control.

We demonstrate that miR-125b, a key node in this microRNA regulatory network, is upregulated in gastric cancer (GC) and associated with poor overall survival through an integrated analysis of microRNA and mRNA profiling of GC revealed a mRNA-regulatory network.So we have employed whole genome microarray expression profiling as a discovery platform to compare the transcriptome profiling of human gastric cells (MKN-45) after 48 hours post-transfection of miR-125b mimic (50nM) and mimic control.Pathway analysis shows that the predicted targets of miR-125b are highly involved in apoptosis/program death pathway，and the robust apoptosis genes, BIK and CASP6 are validated as the directed targets of miR-125b. Gene expression profiling was examined by the Agilent platform (AgilentG3-_GX_1color) with MKN-45 cells and analysis with One-Color Microarray-Based Gene Expression Analysis protocol (Agilent). At 48 hours post-transfection of miR-125b mimic and mimic control, total RNAs were isolated from cells by Trizol reagents. The integrity of total RNAs was evaluated by an Agilent 2100 Bioanalyzer (Agilent). Cy3-labeled target cRNA was prepared by Low Input Quick Amp Labeling Kit (Agilent) according to the manufacturer’s instructions. Labeled cRNAs were hybridized with a SurePrint G3 Human GE 8×60K Microarrays (Agilent).