We then examined the content of D-loop region amplified by P4 primer set in HCC cells with varied ASPH expression level and found that ASPH overexpression greatly impaired mtTFA binding to D-loop region in HepG2 cells (HepG2-ASPH vs HepG2-vec: 6.2±2.9 vs 26.1±8.0, P<0.01), while ASPH silencing facilitated mtTFA binding in MHCC-97 L cells (MHCC-siASPH vs MHCC-vec: 13.9±2.2 vs 7.3±1.8, P<0.01; Figure 6e). This evidence concerns the gene ASPH and hepatocellular carcinoma.