Tumor promoting effect of long non-coding RNA RP11-556E13.1 and its clinical significance as a prognostic factor in hepatocellular carcinoma.

Objective: The aim of this study was to reveal the function of the long non-coding RNA (lncRNA) RP11-556E13.1 (RP11) and its clinical significance in hepatocellular carcinoma. Methods: LncRNA and mRNA expression profiling was performed using lncRNA and mRNA microarrays in 5 pairs of HCC and adjacent tissues. 112 paired HCC and adjacent tissue samples were analyzed by semiquantitative real-time polymerase chain reaction (sqRT-PCR) to evaluate the expression of RP11 and its possible associated clinical significance. Smart silencer RNA (siRNA) was used to knockdown the expression of RP11 in HCC cells. After knockdown, the function of RP11 was determined in vitro using some kinds of cell functional experiments in HCC cells including cell proliferation assay, cell apoptosis assay and cell cycle assay. Western blotting (WB) was performed to detect proteins that were presumably associated with these functional changes. An Affymetrix Human HTA2.0 microarray was used to detect differentially expressed genes in HCC cells after RP11 knockdown. GO enrichment and KEGG pathway enrichment analysis were then performed to elucidate the biological roles of these differentially expressed mRNAs. Total RNA was extracted from Huh7 and Hep3B cells transfected with siRNA or negative control after 48 hours. 3 sample pairs of Huh7 and Hep3B cells were used for the following experiment. 3BRP: Hep3B cells transfected with siRNA; 3BNC: Hep3B cells transfected with negative control RNA.Hu7RP: Huh7 cells transfected with siRNA; Hu7NC: Huh7 cells transfected with negative control RNA;