T-CELL SUBSETS OF PERIPHERAL BLOOD AND SPLEEN IN HEPATOCARCINOMA-BEARING RATS

T-cell subsets from peripheral blood and spleen were investigated prospectively by flow cytometry in a murine intrahepatic implanted tumor model. Peripheral blood OX-19+ cells (pan T-cells) and W3/25+ cells (Th cells) decreased with tumor development in tumor-bearing rats, giving significant differences when compared with healthy rats. No significant changes were found in OX-8+ cells (Ts cells). Hence, Th Ts ratio in tumor-bearing rats was lower than that in control group. Meanwhile, on day 17 following tumor implantation, in tumor-bearing rats with intrahepatic or peritoneal metastasis, there were less W3/25+ cells and more OX-8+ cells than those of hepatoma-bearing rats without metastasis (p<0.01). In the spleen, the phenotypes of lymphocyte markers showed less OX-8+ cells and more OX-12+ cells (B cells) tumor-bearing rats. It was suggested that, in tumor-bearing host, not only was T-cell immune function inhibited, but also there was abnormal distribution of T-cell subsets. It seems that there is a close relationship between tumor growth and its spread and a lower Th/Ts ratio.

Anti-tumor activities of macromolecular fractions of fresh gecko vivo and their induction of Bel-7402 cell differentiation

Objective:To investigate the anti-tumor effect of macromolecular fractions of fresh gecko (M-AG) in vivo and their differentiation-inducing activity in Bel-7402 cells in vitro.Methods:An H22 hepatocarcinoma-bearing mouse model was used to evaluate the anti-tumor activity of M-AG samples.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was applied to analyze cell viability.Cell morphology was observed by phase contrast microscopy.The quantity of the alpha-fetoprotein was detected by a radioimmunoassay.Chromatometry was used to assay the albumin quantity.Activities of alkaline phosphatase and γ-glutamyl trans-peptidase were measured by biochemical methods.Finally,western blotting was applied to assess proteins in the mitogen-activated protein kinase (MAPK) signaling pathway.Results:Macromolecular fractions of fresh gecko exerted a significant anti-tumor effect in mice.The inhibition rate of tumor growth was 63% in the moderate M-AG dose group.Cells treated with M-AG displayed a differentiated state.The treatment lowered alphafetoprotein secretion and significantly decreased the activities of γ-glutamyl trans-peptidase and alkaline phosphatase in Bel-7402 cells.In contrast,M-AG increased the amount of albumin in the cell culture medium.All biochemical indices demonstrated that M-AG induced Bel7402 cell differentiation.Western blotting showed no changes in the quantities of extracellular signal-regulated kinase (ERK) 1/2,p38MAPK,or c-Jun N-terminal protein kinase 1/2.However,M-AG significantly activated the phosphorylation of ERK1/2 in a dose-dependent manner.In addition,M-AG had no significant influence on the expression of nuclear factor-kappa B.Conclusion:Macromolecular fractions of fresh gecko has an anti-tumor activity in H22 hepatocarcinoma-bearing mice in vivo and inhibits Bel-7402 cell proliferation in vitro by inducing cell differentiation related to activation of ERK1/2.