Effect And Mechanism Of Xiao’ai Jiedu Prescription On H22Tumor-bearing Mice And Human Hepatocacinoma SMMC-7721Cells

Objective:Found out more signaling pathways and cytokines associated with Xiao’ai Jiedu Prescription (XJP) through detection of tumor gene expression profiles of H22tumor-bearing mice and further explore the antitumor mechanism of XJP by cell experiment from the cellular and molecular level.Methods:In animal experiments, H22tumor-bearing mice were divided into normal control group,low-dosage XJR group,medium-dosage XJR group,high-dosage XJR group,and Cisplatin group.The differences of gene expression of tumor tissues were discovered by using gene chip technology.The antitumor mechanism of signaling pathways and gene expression were found out by using Gene Ontology (GO) and pathway analysis. The expression of MMP9,MMP12,TIMP2,VEGF and TGF-β gene was detected with real-time quantitative PCR (Realtime-PCR).In cell experiment, human hepatoma SMMC-7721cells were cultured together with pharmacological serum of XJP. The effect of XJP on inhibiting the growth of SMMC-7721was evaluated by methyl thiazolyl tetrazolium (MTT) assay. The morphology of apoptotic cells was observed through electron microscope and the apoptosis rate was detected with Annexin V-FITC by flowcytometry. The expression of TLR2,TRAF6,TGFp, VEGF, MMP2, Survivin and Livin protein was detected with using Western Blot.Results:XJP could change expression of multiple genes in H22tumor-bearing mice, and also significantly affect multiple signaling pathways of tumor growth, apoptosis and immune. XJP could decrease the expression of MMP9,MMP12,VEGF and TGFβ, but increase the expression of TIMP2and IRF1.Pharmacological serum of XJP could significantly inhibite the proliferation of human hepatoma SMMC-7721cells with the best efficiency in24hours.The inhibition rates were4.2%(low-dosage XJR group),12.1%(medium-dosage XJR group),35.8%(high-dosage XJR group), and35.3%(Cisplatin group),and the effect of high-dosage XJR group was similar to that of Cisplatin group(P>0.05). Pharmacological serum of XJP can significantly promote the apoptosis of human hepatoma SMMC-7721cells. Cells showed significant apoptosis morphological changes under electron microscopy. The apoptosis rates were detected after pharmacological serum of XJP acting on human hepatoma SMMC-7721cells for24h with Annexin V-FITC by flowcytometry, and the effect of high-dosage XJR group (23.0±1.6%) was similar to that of Cisplatin group (22.8±4.4%)(P>0.05). As for TLR2,TRAF6,TGF-P,VEGF,MMP2,Survivin and Livin protein expressing contents, the groups of XJP were significantly lower than the normal control group.Conclusion:XJP could inhibit the tumors’growth of H22tumor-bearing mice, and affect tumor proliferation, apoptosis and immune-related genes and signaling pathways.It could decrease the expression of MMP9, MMP12, VEGF and TGF-β of H22tumor-bearing mice, but increase the expression of TIMP2and IRF1. Pharmacological serum of XJP could inhibit the proliferation of human hepatoma SMMC-7721cells and induce the apoptosis.It could reduce the expression of TLR2, TRAF6, TGF-β, VEGF, MMP2and Survivin and Livin protein. All of these may be the anti-tumor mechanism of the XJP.