CRC and paired non-cancer mucosa tissues from three sets of cases were homogenized using the TissueLyser II (Qiagen GmbH), and total RNA was isolated using TRIzol (Invitrogen; Thermo Fisher Scientific, Inc.) and purified with the RNeasy mini kit (Qiagen) according to the manufacturer's instructions. Microarray hybridization and data analysis were performed by KangChen BioTech Co., Ltd., according to the protocols for the miRCURY miRNA Array (v.18.0, Exiqon) which comprised >3,100 probes for capturing miRNAs. Samples were labeled with Hy3 using the miRCURY Array Power Labeling kit (Exiqon). rRNA was removed using the rRNA removal kit, and the remaining mRNA was hybridized to each array in a hybridization oven at 55°C and 20 rpm for 20 h. The array was washed with the Gene Expression Wash Buffer (Exiqon) and scanned using the Axon Genepix 4000B Scanner (Exiqon). Then scanned images were analyzed with the GenePix Pro 6.0 software (Axon Instruments; Molecular Devices, LLC). Background subtraction was performed, and the raw data were normalized using the quantile algorithm. Aberrantly expressed miRNAs were classified as those with a 2-fold change in expression and a P<0.05. The obtained microarray data were deposited in the Gene Expression Omnibus database (accession no. GSE101502).

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