Figure 2: Induction of lncRNAs UCA1 expression by HBx.

(a) The levels of HBx mRNA and UCA1 in LO2-HBx stable cells measured by conventional RT-PCR. (b) The levels of HBx mRNA and UCA1 detected in 10 liver cell lines by RT-PCR. (c,d) UCA1 levels in LO2 cells transiently transfected with an HBx-expressing plasmid pcDNA4.0-HBx at different doses. UCA1 was examined either by conventional RT-PCR (c) or real-time PCR (d). (e) The relative levels of HBx mRNA and UCA1 in Hep3B cells after transfected with different dose of an HBx-expressing plasmid. The value of control was set as 1. (f) The relative levels of HBx mRNA and UCA1 in LO2-HBx and Hep3B cells after transfected transiently with HBx-siRNA or negative control siRNA. β-actin was used as internal control in above experiments. Statistically significant differences are reported (Student t-test) for three independent experiments. ^*P < 0.05; ^**P < 0.01. (g) UCA1 levels measured by real-time PCR in 60 paired tumour and adjacent nontumor tissues. The HCC specimens are divided into two sub-groups based on their relative HBx mRNA status (HBx negative and positive group). Horizontal lines in the box plots represent the median; the boxes represent the interquartile range, and whiskers represent the 2.5th and 97.5th percentiles. Wilcoxon’s signed-rank test was used in this study, P = 0.0244. (h) The inverse correlation of levels of HBx mRNA and UCA1 in HBx positive samples. The ΔCt values (normalized to β-actin) were subjected to Pearson’s correlation analysis (R² = 0.1276, P = 0.0237).