Figure 1

COX-2 is upregulated in HBV-positive cells. (A) HepG2 or HepG2.2.15 cells were serum-starved for 24 hours, after which COX-2 mRNA and protein were detected by qRT-PCR and western blot respectively. (B) COX-2 promoter activity was measured by transfecting HepG2 cells with pCOX-2-Luc and pHBV-1.2 or pHBV-1.2*7, an HBx-deficient HBV mutant. A Renilla luciferase reporter vector pRL-TK was used as internal control. Luciferase activity in each sample was measured at 48 hours post-transfection. All data were normalized to the Renilla luciferase value, and are expressed as mean ± SD, n = 3 (*P < 0.05). (C) HepG2 cells were cotransfected with different amounts of pCMV-HBx, and with pCOX-2-Luc and pRL-TK. Relative luciferase activities were measured at 48 hours post-transfection. Results are presented as fold induction over values obtained in cells not transfected with HBx, and as mean ± SD, n = 3. (D) HepG2 cells were transfected with pCMV-HBx. 48 hours after transfection COX-2 mRNA and protein were determined by qRT-PCR and western blot respectively. (E) PGE2 levels were detected in HepG2.2.15 or HepG2 cells transfected with pCMV-HBx compared with mock transfected cells. Cells were treated with the COX-2 inhibitor NS-398 at 100 μM for 24 hours where indicated. Results are presented as mean ± SD, n = 3 (*P < 0.05). (F) The PBMC lysates from healthy individuals (H1 to H4) or HBV patients (P1 to P4) were used to extract total RNA, COX-2 mRNA was then detected by qRT-PCR.