Research On The Antitumour Compounds Of Anti-Liver-Cancer And Strengthening Body Resistance Decoction By Screening The Whole And Decomposed Recipes In Vivo And In Vitro

Objective:To observe the effects of ALC and its decomposed recipes on proliferation of human hepatoma cell line SMMC-7721 in vitro and transplanted H22 hepatoma in mice modles in vivo to investigate the mechanism.Methods:The inhibition of proliferation in SMMC-7721 cell was assayed by MTT-test.Morphologic changes of apoptosis were observed by Hoechst 33258 staining under fluoroscope.The expressions of cell cycle proteins and apoptosis-associated proteins were detected by flow-cytometric analysis.The effect of ALC on the growth of H22 cells transplanted mice was assessed by analysis of tumour expansion and the inhibition rate of the ALC in vivo. The immunocompetence of mice was assessed by haematological examination and blood chemistry assay.The mechanism of inhibiting tumor was assessed by determination of TNF-αand telomere enzymatic activity in tumour homogenate by ELISA.Results:ALC and its decomposed recipes could inhibit the proliferation of SMMC-7721 cells and the inhibition depended on the dose in vitro.The cells showed morphologic changes of the early stage of apoptosis under the the shrunk round cells,condensed cytoplasma and pycnosis of nucleus.Apoptotic cells and apoptotic body could be found under the microscope and fluoroscope.The analysis of cell cycle indicated that ALC and its decomposed recipes blocked cells at G₀/G₁ phases and the cells of S and G₂/M phase reduced which confirmed the induction of apoptosis by ALC.When treated with ALC and its decomposed recipes for 24 hours,mutant type p53 and bcl-2 were down-regulated.In vivo study showed a significant inhibition on the growth of H22 transplanted tumor in the ALC and its decomposed recipes treated mice groups with negligible damage towards the immunity system compared with the control group.Meanwhile,the expressions of TNF-α were up-regulated and telomere enzymatic activity was decreased in the H22 transplanted tumour.Conclusion:ALC and its decomposed recipes could inhibit the proliferation of the hepatoma cell line SMMC-7721.Cells were blocked at G₀/G₁ phases.The apoptosis was related to the down-regulation of mutant type p53 and bcl-2.ALC and its decomposed recipes could induce cell apoptosis through the p53 and bcl-2 down-regulated.The anti-tumor efficacy of ALC and its decomposed recipes in vivo demonstrated that the whole and decomposed recipes could inhibit the growth of H22 transplanted tumor in mice with negligible damage towards the immunity system via the up-regulated the expressions of TNF-αand decreased telomere enzymatic activity in tumour.