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The in vivo antitumor effects of Icariin and its mechanisms
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Abstract:
To investigate the in vivo antitumor effects of Icarrin (ICA) on H22bearing mice and the related antitumor mechanisms. Methods: C57BL/6 mice were subcutaneously inoculated with H22 hepatocarcinoma cell line to establish H22bearing mice model. 24 h after the inoculation, H22bearing mice were treated with different doses of ICA (9.0 mg/ml, 4.5 mg/ml, and 2.25 mg/ml) and cyclophosphamide (30 mg/kg). When the tumors in H22bearing mice grew to 1 g, the mice were weighed, sacrificed and their spleens and tumors were harvested. The mice sera were also collected for determination of TNFα. Half of the spleens and tumors were made into singlecell suspensions and the other half was fixed by 10% formaldehyde solution for pathology analysis and TUNEL detection. Flow cytometry technique was used to detect the expression of surface antigens CD3, CD4, CD8, and DX5 in the spleen. Annexin V/PI double staining method was used to analyze the early and late apoptosis of tumor singlecell suspension. 1.0×106 cells were used to analyze the cell cycle of the solid tumor cells by flow cytometry. Results: The tumor inhibition rate of cyclophosphamide was 67.16%(P<0.001), and those of high, medium and lowdose ICA groups were 24.63% （P<0.05）, 32.84% （P<0.01）and 5.22%, respectively. Annexin V/PI double staining results indicated that the early apoptosis rate of tumor cells in cyclophosphamide group and mediumdose ICA group were higher than those in H22bearing group \[Cy (911±1.584)%, ICA (7.14±1.376)%, H22bearing group (3.6±2.38)%\]. TUNEL result showed that apoptosis index in ICAmedium group was significantly higher than that in H22bearing group\[(5.35±0.73)% vs (1.03±117)%, P<0.01\]. No TNFα was detected in normal control mice and only very low level of TNFα was found in ICA treatment group and Cy group. Pathology study showed that there were severe necrosis in tumors of medium and highdose ICA groups, with the necrosis found in the capsule and the margin of tumors. A small number of inflammatory cells (lymphocytes and monocytes) were observed in the capsule of tumor and tumor tissues.Conclusion: ICA can effectively inhibit tumor growth, which is probably through inducing early apoptosis of solid tumor cells and the subsequent tumor tissues necrosis.
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