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Fig. 4 | Clinical Epigenetics

Fig. 4

From: Epigenetic dysregulation-mediated COL12A1 upregulation predicts worse outcome in intrahepatic cholangiocarcinoma patients

Fig. 4

MiR-424-5p downregulation contributes to COL12A1 upregulation in clinical iCCA. A The methylation degree of COL12A1 enhancer/promoter region in iCCA and nontumor samples from TCGA. ns, p ≥ 0.05. *p < 0.05, using two-tailed paired Student's t test. B The methylation degree of COL12A1 enhancer/promoter region in iCCA, intraductal tubulopapillary neoplasm of biliary duct (ITPN), intraductal papillary neoplasm of biliary duct (IPNB) and nontumor samples from GSE156299. ns, p ≥ 0.05. *p < 0.05, using two-tailed unpaired Student's t test. C The expression patterns of candidate miRNAs (MiR-424-5p, -424-3p, -497-5p, -15a-5p, -15b-5p, -195-5p and miR-16-5p) in 8 pairs of iCCA samples from TCGA. ns, p ≥ 0.05. *p < 0.05, using two-tailed paired Student's t test. D COL12A1 in HuCCT1 and RBE cells after transfection of miR-NC, miR-424-5p, miR-424-3p, miR-497-5p, or miR-195-5p for 48 h was assessed by immunoblotting, respectively. E The putative binding sites of miR-424-5p in wild-type COL12A1 3'UTR aligned with vertical lines. The mutated bases are presented in red. F Luciferase report activity in HEK-293 T cells after co-transfected with COL12A1 3'UTR report and miR-424-5p mimic for 48 h compared to the control. ns, p ≥ 0.05. **p < 0.01, using two-tailed unpaired Student's t test. G COL12A1 mRNA levels in HuCCT1 or RBE cells after transfection of miR-NC or miR-424-5p for 48 h were assessed by RT-PCR, respectively. ****p < 0.0001, using two-tailed unpaired Student's t test. H MiR-424-5p expression level in 10 pairs of iCCA tissue samples was determined by RT-PCR. **p < 0.01, using two-tailed paired Student's t test. The relationship between COL12A1 mRNA and miR-424-5p in 10 pairs of iCCA samples was assessed by Pearson correlation test. Experiments were triplicated. iCCA intrahepatic cholangiocarcinoma, RT-PCR real-time PCR, TCGA The Cancer Genome Atlas

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