The Research On The Role Of Wnt/β-catenin Signal Transduction Pathway In Ginseng Polysaccharide Induced Apoptosis In Human Nasopharyngeal Cancer Cells

Objeetive：To observe the proliferation and apoptosis effect of ginsengpolysaccharide on human nasopharyngeal cancer cells CNE-2and thepossible mechanism.Methods：To detect the cell proliferation of GPS roled on CNE-2cellsby CCK8assay. CNE-2cells with logarithmic phase were incubated withdifferent concentrations(0,0.1，0.2，0.3，0.4g L-1) of GPS for48h, Flowcytometry was used to demonstrate apoptotic changes in GPS treated cells.Hoechst-33258cell staining and transmission electron microscope wereused to demonstrate cell morphology changes. The expression of β-cateninmRNA was identified by Real-time PCR. The localization and expression ofβ-catenin and TCF4were detected by the immunofluorescence staining withlaser scanning confocal microscope (LSCM). The expression of β-catenin,Bcl-2and Bax proteins is detected by western blot.Results: CCK8results showed that GPS can inhibit the proliferation ofCNE-2cells in dose-and time-dependent manner, IC50of GPS in48h was0.39g·L-1to CNE-2cells. When treated with0.1，0.2，0.3，0.4g L-1of GPS for48h, the cells showed cell apoptotic rates of5.69±0.29%,10.3±0.63%,15.4±0.74%and35.7±1.86%respectively,statistical significance could be found compared with the controlgroup(2.08%±0.11%)(P<0.05). Hoechst33258staining images showedapoptotic morphology change and apoptotic body could be found in CNE-2cells induced by GPS conditioned medium at0.4g L-1for48h. The results ofReal-time PCR showed the level of mRNA of β-catenin was decreased. Theimmunofluorescence staining results showed that the expressions ofβ-catenin and TCF4decreased significantly in the nucleus and β-cateninprotein positive particles transferred from the nucleus to the cytoplasm.Western blot showed the expression of proapoptosis protein Bax enhanced,antiapoptosis protein Bcl-2protein and β-catenin decreased significantlywith an increasing density GPS for48h (P<0.05).Conclusion: GPS significantly inhibited the proliferation of CNE-2cells and promoted apoptosis. GPS inhibited Wnt/β-catenin signalingpathway which may be the important mechanism that GPS inducedapoptosis of human nasopharyngeal cancer CNE-2cells.