Experimental Study On Anti - Tumor Invasion And Metastasis Of Ginsenoside Structural Modified

ObjectiveTo investigate the effects and mechanism of HRG, a ginsenoside structure modification on the proliferation, growth, cell cycle distribution, ability of invasion and metastasis of B16-F10mouse melanoma cells.Methods(1) B16-F10mouse melanoma cells were cultured in vitro with different concentrations of HRG(12.5、25、50、100μg·ml-1). The cell proliferation inhibiting rate after HRG affected cells24,48,72hours was detected by MTT assay; the cell growth curve was detected by cell count assay and the cell cycle distribution was examined by flow cytometer.(2) The cell migration and invasion were detected by migration assay and transwell assay in vitro, respectively; the effects of HRG on the expression of migration-related proteins (matrix metalloproteinase-2, matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1and tissue inhibitor of metalloproteinase-2) were detected by Western blotting.(3) Mouse spontaneous melanoma model and artificial melanoma model were established in vivo to observe the effects of HRG on the growth and tumor metastasis. In these models, mice were randomly divided into five groups, i.e., the model control group, the Rg3control group, the small, medium, large dose HRG groups. Normal saline whose mass fraction is0.9%were given to mice in the model control groups, Rg3were given to mice in the Rg3control groups at the daily dose of20mg· kg-1by peritoneal injection. HRG was given to mice in the the small, medium, large dose HRG groups at the daily dose of10,20and40mg· kg-1by peritoneal injection.Results(1) B16-F10mouse melanoma cells in each HRG group and Rg3group could inhibit the proliferation of B16-F10mouse melanoma cells compared with the blank control group in a time and dose dependent manner; as the drug concentrations increased, inhibition of HRG on B16-F10mouse melanoma cells enhanced with time-effect dependent and dose-effect dependent; the effect on cell cycle showed that HRG could regulate the cell cycle. B16-F10mouse melanoma cells in S phase were blocked significantly and the number of cells in divisions decreased. There were significant differences between HRG groups and the blank control group (P<0.01).(2) After the treatment of50μg· ml-1and100μg·ml-1HRG, the number of B16-F10mouse melanoma cells through the transwell chambers were significantly decreased in a dose-dependent manner, there were significant differences between HRG (50、100μg·ml-1) groups and the blank control group(P<0.001).The expression of migration-related proteins (matrix metalloproteinase-2, matrix metalloproteinase-9) were reduced by HRG in a dose-dependent manner while expressions of tissue inhibitor of metalloproteinase-1and tissue inhibitor of metalloproteinase-2were not changed(P>0.05).(3) In mouse spontaneous melanoma model, these were no significant difference in primary foci in each HRG group, Rg3control group and model control group (P>0.05). The number of lung metastatic nodes significantly decreased in each HRG group and Rg3control group (P<0.01). The inhibition rate of tumor metastasis were22.22%,43.62%,57.20%and33.74%, respectively.In mouse artificial melanoma model, the number of lung metastatic nodes significantly decreased in each HRG group and Rg3control group(P<0.01). The inhibition rate of tumor metastasis were17.08%,47.69%,55.16%and32.03%, respectively.In mouse spontaneous melanoma model and artificial melanoma model,20,40mg·kg-1HRG could advance the thymus index of the tumor-bearing mice while there was no impact on lienal index(P>0.05).Conclusion(1) HRG inhibited the proliferation of B16-F10mouse melanoma cells in a time and dose dependent manner as well as regulating cell cycle and thus inhibited cell proliferation.(2) HRG could inhibit the invasion and metastasis of B16-F10mouse melanoma cells in vitro and in vivo, and the mechanism might relate with down-regulating the expression of MMP-2and MMP-9proteins.(3) HRG could inhibit lung metastasis of B16-F10mouse melanoma and improve immune function. Nevertheless, there was no effect on carcinoma in situ.