methylation and expression. a Promoter methylation status of the study patient’s samples was examined. Methylation-specific PCR (MSP) and unmethylation-specific PCR (UMSP) were performed. Only tumor tissue had promoter methylation. b Promoter methylation status of the COL1A1 gene six hepatcellular carcinoma (HCC) cell lines. Complete methylation was detected in the cell line HLE; partial methylation in HLF, HuH2, and SK-Hep1; and complete unmethylation in Hep3B and HuH7. c
COL1A1 expression was reactivated in HLE, HLF, HuH2, and SK-Hep1 by 5-aza-2′-deoxycytidine (5-aza-dC) treatment. β-Actin was used as the normalization gene. d COL1A1 protein expression was confirmed by western blotting. Very weak or no band was detected in the cell lines with positive promoter methylation. β-Actin was used as the normalization gene.