Hepatitis B virus X gene induces human telomerase reverse transcriptase mRNA expression in cultured normal human cholangiocytes

Figure 1 Transfection efficiency evaluated by EGFP-express-ing cells in the HBx gene transfected cell cultures and empty vector control. pEGFP 0.1 μg and pCMV-X (or pcDNA3) 2.9 μg were co-transfected into HBECs by Lipofectamine. Thirty-eight hours after transfection. EGFP-expressing cells were visible under fluorescence microscope in the HBx gene transfected cell cultures (A) and empty vector control (B), but not in the blank control (Under fluorescence microscope × 200).

Figure 2 Analysis of hTERT mRNA expression by RT-PCR. RT-PCR was performed on total RNA extracted from HBECs transfected with OPTI-MEN medium (1), pCMV-X (2) and pcDNA3 (3), respectively. M: DL2000 Marker.

Figure 3 Quantitative and relative changes of hTERT mRNA expression analyzed by NIH Image software. Dramatic expres-sion of hTERT mRNA was observed in HBECs when trans-ferred with pCMV- X vector (lane 2), but there was no hTERT mRNA expression in HBECs when transferred with OPTI-MEM medium (lane 1) and empty vector (lane 3).

Figure 4 HBx protein expression in transferred HBECs assayed by ultrasensitive immunocytochemistry. The blank vector and OPTI-MEN transfected HBECs showed no expression of HBx protein (A, C), but pale brown positive signals scattered in pCMV- X vector transferred HBECs (B). Immunocytochemis-try (S-P methods, × 200).