The Research Of Ginsenoside Rg3 Enhances The Effect Of Sensitized DC-induced CTL On Mouse Kidney Cancer Cell RAG

Ginsenoside Rg3 is an effective component of ginseng(Panax ginseng C.A.Meyer).Recent studies have confirmed that Rg3 can induce tumor cell apoptosis,selectively inhibit tumor cell adhesion and infiltration,and inhibit tumor neovascularization.At the same time,it can regulate the body's immune function and enhance the body's non-specific immune function.It is considered to be an immune enhancer.Dendritic cells(dendritic cells,DC)are a very powerful antigen-presenting cell and play an important role in the immune response.When an immune reaction occurs in the body,DC can efficiently promote the body to generate cytotoxic T lymphocytes and helper T cells,and generate an immune response.DC is not only the initiator of the body's immune response,but also a participant.The CTL-mediated cellular immune response activated by DC plays an extremely important role in the occurrence,development and prognosis of malignant tumors.Kidney cancer is one of the common tumors in the urinary system.Like most tumors,the treatment effect is not satisfactory,and most patients die of tumor metastasis.Therefore,the research on the migration and metastasis of kidney cancer has always been one of the hot topics discussed by scholars.In this study,Rg3 was used to enhance RAG-sensitized DC-induced CTL in mouse kidney cancer cells,and Rg3-DC vaccine was prepared.The immune enhancement effect of Rg3 on DC and the anti-tumor mechanism of Rg3 have multiple anti-targets,multiple links,and multiple effects.The characteristics of RAG cells cultured in vitro in mouse kidney cancer were studied and compared.The effects of Rg3-DC vaccine and conventional DC vaccine on RAG cell proliferation,migration,killing and immunological function were not reported.At the same time,it further explores the anti-tumor mechanism of Rg3-induced DC,and lays a theoretical foundation for the preparation of a new and potent DC vaccine.Object:1.To explore the role of Rg3 in boosting DC immunity and compare the effects of Rg3 DC vaccine and conventional DC vaccine on T cells;2.To study the effects of Rg3 on RAG-sensitized DC-induced CTL on mouse kidney cancer RAG,and to investigate the effects of Rg3 on tumor suppression,invasion,metastasis and immunological function,and to explore the anti-tumor mechanism of Rg3 on DC immune function.Method:1.The study of Rg3 activating DC function,using FCM to detect the expression of DC surface molecules(CD11c PE,CD80,CD86,MHC?),and observe the effect of Rg3 on DC maturity after 5 days of treatment with different concentrations of Rg3(2,10,20);2.Rg3 enhanced RAG-sensitized DC corresponds to different concentrations of heterogeneous T cells(DC: T : 1: 100,1:10,1: 1)mixed culture for 48 h,using CCK8 method to detect T cell proliferation rate,ELISA Immune Function;3.Incubate the induced CTL cells with RAG(CTL: RAG effect target ratio: 10: 1,20: 1,40: 1),after 18 h,use CCK8 to detect the killing rate,FCM to detect the apoptosis;Western blot to detect the apoptosis Gene(Bax,Bcl-2);4 Transwell and scratch healing experiments proved the effect of Rg3-DC + T on migration and metastasis of RAG cells.Result:1.Ginsenoside Rg3 can promote DC maturation;2.FCM test results showed that with the increase of Rg3 concentration,the induced DC expression levels of CD86,MHC?,and CD11 c increased,and increased with time,while CD80 did not increase.3.CCK8 test results showed that Rg3-DC induced CTL cells were mixed with RAG.With the increase of RAG ratio of tumor cells,CTL had a greater effect on tumor cells.The lethal effect gradually decreases,and the apoptosis rate detected by FCM decreases;4.Transwell and scratch healing experiments show that Rg3-DC-induced CTL cells have a significant effect on RAG cells cultured in vitro.Conclusion:(1)Ginsenoside Rg3 can promote the differentiation and maturation of dendritic cells and stimulate the proliferation of lymphocytes,and has a certain dose-effect relationship;(2)The value-added effect of antigen-sensitized Rg3-DC on CTL cells shows that a certain concentration of Rg3 can optimize DC to enhance its ability to capture antigens and activate CTL.(3)The CTL cells in this study have an inhibitory effect on the growth,invasion and metastasis of RAG.