ObjectiveTo evaluate the effect of pre-infusion of allogeneic lymphoyctes treated with 5-FU on the rat liver graft. MethodsRat liver transplant models from Wistar to SD were established. Four groups were designed as following: control group: only liver transplantation without any other intervention; lymphocytes group: 1 ml of untreated lymphocytes (5×106/ml) from Wistar rats were preinfused into SD rats on day 7 and 4 separately before transplantation; lymphocytes with low concentration of 5-FU group: low concentration 5-FU (7.5 μg) treated lymphocytes were preinfused as above; lymphocytes with high concentration of 5-FU group: high concentration 5-FU (15 μg) treated lymphocytes were preinfused as above. Fas-L and CD8 expression were detected by immunohistochemistry method on day 7 after transplantation. ResultsThe integral opticaldensity (IOD) of Fas-L positive lymphocytes in the lobules of liver and portal areas were higher in lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. There was no difference between lymphocyte group and lymphocytes with high concentration of 5-FU group （Pgt;0.05）. The IOD of CD8+ expression in lobules of liver was not different among all the three lymphocytes treated groups （Pgt;0.05）. But in portal areas, CD8+ expression was lower in the lymphocytes with low concentration of 5-FU group than in the other groups （Plt;0.05）. ConclusionPreinfusion of lymphocytes treated with low concentration 5-FU can induce graft immune tolerance, the probable mecanism of which is the increasing Fas-L expression in graft.
ObjectiveTo evaluate therapeutic effect of the targeting recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3 on primary hepatocellular carcinoma cells (HepG2) and subcutaneous implanted tumors of nude mice. MethodsHepG 2 cells, breast cancer MDA-MB-231 cells， and normal hepatic L-02 cells were infected with the previously constructed targeting recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3, then morphological change was observed and apoptosis index was detected by cell morphologic assay. Tumor growth and pathological changes were observed after the implanted tumors model of nude mice were injected with the recombinant adenovirus. Results Both apoptosis and obvious apoptotic peak of cells were observed by flow cytometry (FCM) in HepG 2 cell group. The apoptosis index of MDA-MB-231 and L-02 cells was 0 and 7.3%, respectively, which were significantly lower than that of HepG2 cells (48.2%), Plt;0.01. The volume of subcutaneous tumor of nude mice was (0.26±0.31) cm 3 in HepG2 cell group, (0.25±0.15) cm 3 in MDA-MB-231 cell group， and (0.26±0.28) cm 3 in control group on two weeks after implantation of cancer cells, and no statistical difference was found among them (Pgt;0.05). The volume of subcutaneous tumor of nude mice was (0.53±0.12) cm 3 in HepG 2 cell group, (0.49±0.22) cm 3 in MDA-MB-231 cell group， and (0.54±0.13) cm 3 in control group on four weeks after implantation of cancer cells, and the difference was not significant among them (Pgt;0.05). But on eight weeks after implantation of cancer cells, the volume of tumor was (0.65±0.13) cm 3 in HepG 2 cell group, (1.27±0.32) cm 3 in MDA-MB-231 cell group， and (1.43±1.09) cm 3 in control group, which suggested that the growth of tumor in HepG 2 cell group slowed down significantly when compared with MDA-MB-231 cell group and control group (Plt;0.01), although the difference in the latter two groups was not significant (Pgt;0.05). The necrosis regions were found with lymphocytic infiltration and apoptosis cells in HepG2 cell group after injection of recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3, but no pathological change of cells was found in control group after injection. It was found that the cells in MDA-MB-231 cell group were solidly arranged with small glandular lumenlike structure, marked cellular atypia, multinucleated tumor cells and megakaryocytic tumor cells, which was not different from that before injection. ConclusionThe targeting recombinant adenovirus Ad-EAFP-PALB/r-Caspase-3 induces the apoptosis of primary hepatocellular carcinoma in vivo and in vitro, which may provide new ways of targeted gene therapy.