Study On Immunogenicity Death Induced By NsPEFs In Cervical Cancer Cells And Stimulating Immune Response Of Solid Tumors After Ablation

Objective:(1)To investigate the killing effect of nsPEFs on cervical cancer cells of different types and whether it can induce immunogenicity death of cervical cancer cells.(2)To observe the ablation effect of nsPEFs on subcutaneous cervical cancer transplant tumor in mice and the specific immune response.Methods:(1)The suspension of human cervical cancer Hela cells and mouse cervical cancer U14 cells were stimulated by nsPEFs with different electric field strength.The cell viability,apoptosis rate and expression of danger associated molecular patterns(DAMPs)of tumor cells after electric shock were detected by CCK-8 method,flow cytometry and ELISA method respectively.(2)These animal models of subcutaneous transplanted tumor of cervical cancer in mice was constructed.The effect of nsPEFs ablation and the body immune response of cervical cancer soild tumors of mice were detected.The experiment was divided into three groups of blank control group,tumor group and stimulation group.To observe the weight of mice and the growth changes after nsPEFs ablation of solid tumors.Flow cytometry was used to detect the differentiation and maturation of DCs,the differentiation and activation of T cells and the release of cytokines by T cells in the lymph nodes,spleen and solid tumor tissues of mice after the ablation of nsPEFs.Results:(1)With the increase in electric field strength and the prolongation of time,the viability of cervical cancer cells became lower and the apoptosis rate obviously increased after nsPEFs stimulation.The expression of calreticulin(CRT),heat shock protein 70(HSP70)and high mobility group box-1(HMGB1)in Hela or U14 cells was significantly increased at 24 h or 3 h after nsPEFs stimulation above 10 kV/cm respectively.(2)After cervical cancer subcutaneous transplant tumors of mice were ablated by nsPEFs,the ponderal growth of mice slowed,the volume of tumor increased rapidly and increased to the peak after nsPEFs ablation,and the spleen index increased significantly.After nsPEFs ablation of subcutaneous transplant tumor,the differentiation and maturation of DCs,the differentiation and activation of CD8~+and the cytokines of IFN-?,TNF-?and GzB released in CD4~+and CD8~+cells were significantly increased in the stimulation group than in the tumor group in solid tumor tissue.The frequency of DCs and the expression of cytokine TNF-?in lymph nodes and spleen tissues increased significantly in the stimulating group than in the tumor group,while the activation of CD4~+cells decreased significantly.Conclusion:(1)nsPEFs has different killing effects on different types of cervical cancer cells and can induce apoptosis of cervical cancer cells to release DAMPs signals,which can lead to the phenomenon of immunogenic cell death.(2)nsPEFs can effectively ablation the solid tumor transplanted in mice and inhibit the growth of tumor.(3)After cervical cancer subcutaneous transplant tumors of mice were ablated by nsPEFs,the response of local and systemic immune systems was activated by promoting the differentiation and maturation of DCs,the differentiation and activation of T cells and the release of various immune cytokines such as IFN-?,TNF-?and GzB.