关键词: 关键词:SFRP1, 启动子, 荧光素酶报告基因载体, HCV Core

Abstract: Objective To investigate the effects of HCV Core on SFRP1 gene promoter activity. Methods Different lengths of the 5’-flanking region of SFRP1 promoter were amplified from hepatoma cell line Huh7 genome DNA by PCR methods. The products were cloned into pGL3-Basic vector. The recombinant reporter plasmids and control plasmid pRL-TK were cotransfected into HEK293 to determine the strongest activity area of SFRP1 promoter. To examine whether HCV Core could inhibit SFRP1 promoter activity, hepatoma cell line SK-Hep1 were transfected with reporter plasmids and then infected with AdCore or AdGFP control. The promoter activity was measured by luciferase activity. Results The strongest activity area of SFRP1 promoter was -407～-27nt . Compared with pGL3-Basic negative control, the relative luciferase activity increased by 37.31±4.45 folds (P<0.01). And other reporters(pGL3-S2～S5) increased by28.74±2.47, 13.56±2.52, 12.97±0.87and 8.29±0.09 folds respectively(P<0.01). HCV Core protein could inhibit SFRP1 promoter activity. The inhibitory effect on pGL3-S1 was the strongest, with inhibitory rate 63.8%±1.0% (P<0.01). Conclusions HCV Core protein inhibits SFRP1 promoter activity, resulting in down-regulation of SFRP1 mRNA expression level, thus participating in HCC carcinogenesis.

Key words: Key words:SFRP1, promoter, luciferase reporter vector, HCV Core