Fig. 1: HDAC2-AS2 was upregulated by TGFβ.

a Experimental layout to identify TGFβ dependent lncRNAs. HepG2, Huh7, MHCC97, and PVTT cells were treated with TGFβ (2.5 ng/mL) for 8 h. RNA was harvested for microarray analysis of lncRNAs. Then our lncRNA microarray (TGFβ treatment vs. ctrl) merged with upregulated lncRNAs in the TCGA database (HCC samples vs. non-tumor tissues). qRT-PCR validation of indicated 11 lncRNAs relative to β-actin in MHCC97H cells treated with TGFβ (2.5 ng/mL) for 48 h. b qRT-PCR validation of HDAC2-AS2 relative to β-actin in LO2, PVTT, Huh7, SMCC7721, MHCC97L, and MHCC97H cells treated with TGFβ (2.5 ng/mL) for 48 h. c MHCC97H cells were pretreated with 10 μM SB431542 for 1 h, followed by TGFβ treatment for 48 h. HDAC2-AS2 expression was detected by qRT-PCR. d qRT-PCR analysis of HDAC2-AS2 expression relative to β-actin in SMCC7721 WT and Smad4 KO cells treated with TGFβ (2.5 ng/mL) for 48 h. e qRT-PCR analysis of HDAC2-AS2 relative to β-actin in HepG2 and HepG2.2.15 cells. f qRT-PCR analysis of HDAC2-AS2 relative to β-actin in HepG2-NTCP cell line after HBV infection. g qRT-PCR analysis of HDAC2-AS2 expression relative to β-actin in human HCC tissues and paired adjacent noncancerous tissues (n = 10 for paired samples). h Elevated HDAC2-AS2 expression in the tumor of HCC (n = 50, 200 for NT and T samples) in the TCGA database. i Boxplot showing the expression levels of HDAC2-AS2 in tumor (T) and nontumor tissues (NT) sourced from the TCGA-LIHC database, classified according to their etiology (n = 53, 36, 151 for samples of HBV, HCV, and others, respectively). The boxplots denote the median with a quartile range (25–75%), and the length of whiskers represents 1.5× the IQR. j The expresision levels of TGFB1 in HDAC2-AS2 low- (n = 157) and high- (n = 83) expressing tumor and nontumor tissues. The boxplots denote the median with a quartile range (25–75%), and the length of whiskers represents 1.5× the IQR. k GSVA analysis of epithelial-mesenchymal transition, inflammatory response, and lymphocyte-mediated immunity signatures in HDAC2-AS2 high-expressing tumor tissues. The P-values were calculated based on a two-tailed permutation test. NES, normalized enrichment score. Error bars represent mean ± SEM for three independent replicates. Statistical analyses were determined by two-tailed Student’s t-test (b–j). Source data are provided as a Source Data file.