X-ray Induced Pyroptosis Through Caspase 3/GSDME Signaling Pathway To Promote Anti-tumor Immunity

GSDME protein is a key efficent protein in cell pyroptosis,and many studies have shown that GSDME-induced pyroptosis plays an important role in cancer therapy.Pyroptosis can cause local inflammation and attract inflammatory cell infiltration as a highly immunogenic form of cell death,which plays an important role in alleviating immunosuppression of tumor microenvironment and inducing systemic immune response.Cancer is a kind of multi-gene disease with complex etiology.The incidence trend of cancer is getting younger and younger,which is harmful to human health.With the progress of science and technology and the gradual deepening of medical research,the treatment of tumor is becoming more and more perfect,common surgical treatment,chemotherapy,radiotherapy and immunotherapy,among which radiotherapy is still a very important therapy for tumor treatment,intracellular caspase-3 can be activated by radiation to cut and activate GSDME to cause pyrodeath.Promote phagocyte phagocytosis of tumor cells.In this study,the cell model with high expression of GSDME protein was screened,and the changes of phagocytes’ phagocytosis to tumor cells were observed after X-ray treatment,so as to explore the relationship between cell pyroptosis induced by X-ray through caspase3/GSDME signaling pathway and anti-tumor immunity.Objective Selecting tumor cells with high GSDME expression and determining the optimal irradiation conditions for inducing pyroptosis of tumor cells treated with X-ray irradiation.Observing the condition of being devoured of tumor cells before and after treatment,and exploring the relationship between pyroptosis induced by X-ray through caspase3/GSDME signaling pathway and anti-tumor immunity.Methods1.Tumor cells with high expression of GSDME were screened In the experiment,HCT116(human colon cancer cells),Hela(human cervical cancer cells)and Ovcar8(human ovarian cells)were selected to investigate the expression of GSDME protein in different cells by Western blot.2.To determine the optimum conditions for inducing pyrodeath of tumor cells Determining the optimal irradiation conditions for inducing pyroptosis of tumor cells treated with X-ray irradiation.Tumor cells were treated with different X-ray irradiation doses and time,and the optimal irradiation dose and time were selected to induce pyrodeath of tumor cells.3.Tumor cells and phagocytes were cultured together to observe phagocytosis.The tumor cells in treatment group and control group were cultured together with phagocytes respectively,and the changes of tumor cells were observed by fluorescence microscopy.Results:Hela cells showed high expression of GSDME protein through Westernblot analysis,and Hela cells were identified as tumor cells in this study.0Gy,2Gy and 15 Gy dosage groups were set up to observe the pyrodeath of cells at 0h,48 h,72h and 96 h respectively.The results showed that Hela cells incubated for 72 h at the irradiation dose of 8Gy showed obvious pyroptosis,and pyroptosis induced by caspase3/GSDME signaling pathway was the best.Phagocytosis results showed that Hela cells after pyroptosis induced by irradiation had better phagocytosis effect by M1 macrophages,more phagocytosis effect and better anti-tumor immunity effect.Conclusion:1.After being irradiated with 8Gy for 72 h,Hela cells were observed to have obvious GSDME protein expression and obvious pyroptosis.2.Irradiated Hela cells(treated group)and unirradiated Hela cells(control group)were co-cultured with M1 macrophages,and more Hela cells were phagocytosed in the treated group,indicating that X-ray induced pyroptosis of tumor cells through caspase3-GSDME pathway promoted anti-tumor immunity,which could provide a new idea for subsequent cancer treatment.