Profiling mRNA and miRNA Levels

Total RNAs were extracted with Trizol reagent according to the manufacturer’s instruction (Invitrogen, CA, USA). To determine the mRNA levels of AR, YB-1 and PSA, total RNAs were reversely transcribed by iScript cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA), respectively, according to the manufacturer’s instructions34 (link). The 18 S rRNA gene was used as reference for normalization34 (link). Taqman RT-qPCR was performed to detect mature miRNA expression using Taqman miRNA reverse transcription kit, has-miR-190a (AB Assay ID: 000489) and RNU6B (U6, AB Assay ID: 001093) according to the manufacturer’s protocol (Applied Biosystems)35 (link). The U6 was used as reference for normalization35 (link).