Hepatitis B virus X protein suppresses adriamycin-induced apoptosis of hepatocellular carcinoma cells and expression of p53 and PTEN / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of hepatitis B virus X protein (HBx) on adriamycin-induced apoptosis of hepatocellular carcinoma cells and the expressions of p53 and PTEN.</p><p><b>METHODS</b>HepG2, HepG2/GFP, and HepG2/GFP-HBx cells were treated with adriamycin (2.5 microg/ml), and the apoptotic cell death was determined by observing the morphological changes and flow cytometry. The expressions of p53 and PTEN mRNA in the 3 cells were detected by RT-PCR, and the expressions of p53 and PTEN protein were analyzed by Western blotting.</p><p><b>RESULTS</b>Adriamycin induced significant cell death in HepG2 and HepG2/GFP cells, which became rounded, shrunk, and detached after the treatment; but no significant cell death occurred in HepG2/GFP-HBx cells. Flow cytometry analysis showed that the apoptotic rate was significantly lower in HepG2/GFP-HBx cells (3.94%) than in HepG2 (59.03%) and HepG2/GFP cells (61.38%) at 36 h after the treatment (P<0.001), while no significant difference was observed between HepG2/GFP-HBx (3.94%) and the control cells (2.12%, 2.78%, and 2.55%) (P>0.05). RT-PCR showed lowered expression of PTEN mRNA in HepG2/GFP-HBx cells as compared to that in HepG2 and HepG2/GFP cells, while no significant difference was noted in p53 mRNA. Western blot analysis showed that PTEN protein decreased while p53 protein remain unchanged in HepG2/GFP-HBx cells.</p><p><b>CONCLUSION</b>HBx suppresses adriamycin-induced apoptosis of HepG2 cells and PTEN expression. The inhibitory effect of HBx on the cell apoptosis may be related to the inhibition of p53-PTEN pathway.</p>

Hepatitis B virus X protein suppressing adriamycin-induced apoptosis of HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effect of hepatitis B virus X protein (HBx) on adriamycin-induced apoptosis of hepatocellular carcinoma cells.</p><p><b>METHODS</b>HBx gene fragment was amplified from subtype adr HBV plasmid by PCR, and inserted into Hind III and Kpn I sites of green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP/HBx. The pEGFP-C1 and pGFP-HBx were introduced into HepG2 cells by Lipofectamine 2000 to obtain HepG2 cells expressing GFP. GFP-HBx fusion protein was selected using G418. The expression of HBx gene was demonstrated by RT-PCR analysis. HepG2, HepG2/GFP and HepG2/GFP-HBx cells were treated with adriamycin (2.5 microg/ml), and apoptosis of the cells was determined by their morphological changes, trypan blue exclusion, and flow cytometry analysis.</p><p><b>RESULTS</b>Under a fluorescence microscope, visible expression of GFP and GFP-HBx fusion proteins were observed in HepG2/GFP and HepG2/GFP-HBx cells, even after growing over 70 generations. RT-PCR analysis showed that HBx gene was expressed in HepG2/GFP-HBx cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in HepG2 and HepG2/GFP cells while no significant cell death was observed in HepG2/GFP-HBx cells. Flow cytometry analysis showed that apoptosis rates in HepG2/GFP-HBx (3.94%) cells were significantly lower than those in HepG2 (59.03%) and HepG2/GFP cells (61.38%) at 36 hours after the adriamycin treatment (P < 0.01). No significant differences of apoptosis rates of HepG2/GFP-HBx (3.94%) and of the untreated cells (2.12%, 2.78%, 2.55%) (P > 0.05) were observed.</p><p><b>CONCLUSION</b>A HepG2 cell line expressing GFP and GFP-HBx fusion proteins was successfully established. HBV X protein blocks adriamycin-induced apoptosis of these HepG2 cells.</p>

Effects of Chinese herbs for cool-moistening and freeing collaterals on serum gastrin and surface electrogastrogram in patients of diabetes mellitus with gastroparesis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Chinese herbs (CH) for cool-moistening and freeing collaterals on gastro-dynamic disturbance in patients of diabetes mellitus type 2 with gastroparesis (DM-GP).</p><p><b>METHODS</b>Fifty-three patients of DM-GP were enrolled and treated with CH (n = 28) and Cisapride (n = 25) respectively for 4 weeks, the changes of gastrin and electro-gastrogram (EGG) before and after treatment were observed.</p><p><b>RESULTS</b>After treatment, the EGG improved significantly, showing the rhythm significantly improved, and level of serum gastrin lowered significantly, as compared with those before treatment, the difference was significant (P<0.01), but insignificant difference was found between the two groups. Fifteen patients in each group were followed-up afar stopping medication for 3 months, recurrence occurred in 1 patient of CH treated group, and 2 patients of Cisapride treated group. No adverse reaction was found in the rest patients.</p><p><b>CONCLUSION</b>CH could obviously improve the gastro-intestinal motility and hormones abnormality.</p>