RNA-Binding Protein Trx regulates alternative splicing and promotes metastasis of HCC via interacting with LINC00152

RNA-binding proteins (RBPs) were reported to be the key regulator of epithelial-mesenchymal transition (EMT)-related post-transcriptional in tumor diseases.To explore the role of RBPs in metastasis of hepatocellular To explore the role of RBPs in metastasis of hepatocellular carcinoma (HCC), whole transcription sequencing was conducted to identify differential RBPs between HCC with metastasis or without metastasis. The influence of RBPs on metastasis of HCC was verified by in vitro and in vivo experiments. Interaction of RBPs with non-coding RNAs was evaluated by RNA immune-precipitation and pull-down assay. RNA-seq whole genome sequencing and alternative splicing analysis were further performed to clarify post-transcription regulation mechanisms. 3 HCC tissues with metastasis and 3 HCC tissues without metastasis were selected for whole transcript sequencing. Total RNA was extracted and purified with magnetic beads. After fragmentation of 2nd-cycle single-stranded cDNA (ss-cDNA), sample was labeled and hybridized to the Affymetrix Human Gene 1.0ST Array. The data were analyzed with Robust Multichip Analysis (RMA) algorithm using default analysis settings and global scaling as normalization method by Partek® Genomics Suite 6.6. Values presented are log2 RMA signal intensity. Normalized data were further analyzed using one-way ANOVA to screen out the differential expression gene.