The Research Of The Effect And Mechanism Of B1 On Anti-lung Cancer

Background and purposeWith changes in the environment and living habits,the global cancer incidence rate is increasing rapidly.According to the "Global Cancer Report" released by the International Agency for Research on Cancer(IARC),there will be 19,290,000 new cancer cases and about 9,960,000 deaths worldwide in 2020,indicating that the global cancer situation is further serious.Lung cancer is the second most common cancer in the world.In 2020,there will be 1,800,000 lung cancer deaths,ranking first in cancer deaths.However,the currently used chemotherapy drugs such as cisplatin have serious adverse reactions,so it is imminent to find new and effective anti-lung cancer drugs.MethodsThe effects of B1 on cell viability,proliferation,invasion and migration of A549 cells were detected by CCK8,Ki67,Transwell and scratch assays;A tumor model was constructed by subcutaneous injection of human lung cancer A549 cells on the back.BABL/c Nude mice were randomly divided into 5 groups,Model group,DDP group,1,3,10 mg/kg B1 dose groups.Take the material 28 days after the administration,take pictures of the tumor tissue and measure the volume and weight;The tumor tissue were stained with HE to detect the effect of B1 dose groups on the tumor;LDH,Hoechst33342/PI double staining assay and flow cytometry were used to detect the effect of pyroptosis in human lung cancer A549 cells;ELISA kits and immunofluorescence were used to detect the expression of IL-18,IL-1β and Caspase-1 in A549 cells and lung tumor tissues;Western Blot was used to detect the influence of NF-κB,NLRP3,GSDMD,and N-GSDMD protein content in A549 cells and tumor tissues;After administration of Caspase-1 inhibitor VX-765,LDH activity kit,Hoechst33342/PI double staining experiment and flow cytometry were used to detect the effect of pyroptosis in A549 cells;After treatment with the Caspase-1inhibitor VX-765,IF and Western Blot were used to detect the expression of GSDMD and N-GSDMD in A549 cells in each group;After administration of NF-κB inhibitor BAY,LDH activity kit,Hoechst33342/PI double staining assay and flow cytometry were used to detect the effect of pyroptosis in A549 cells;After administration of NF-κB inhibitor BAY,the expressions of NF-κB,NLRP3,GSDMD and N-GSDMD in A549 cells in each group were detected by IF and Western blot.ResultsB1 can significantly reduce the viability,proliferation,invasion and migration of A549 cells;B1 groups can reduce tumor volume and weight,and inhibit the growth of tumor cells and tissues;B1 increased the activity of LDH,the proportion of PI-positive cells,and the ratio of pyroptotic cells in A549 cells,which suggests that B1 promotes pyroptosis;B1 can promote the expression of NF-κ B,NLRP3,Caspase-1,GSDMD and N-GSDMD in A549 cells and tumor tissue,and promote the release of IL-18 and IL-1β;After administration of Caspase-1 inhibitor VX-765 to A549 cells,Characteristics of B1-induced pyroptosis,such as LDH activity,proportion of PI-positive cells and proportion of pyroptotic cells were inhibited;After A549 cells were given the Caspase-1 inhibitor VX-765,the effect of B1 on promoting the expression of Caspase-1,GSDMD and N-GSDMD was significantly inhibited;After A549 cells were given the NF-κB inhibitor BAY,the characteristics of B1-induced pyroptosis,such as LDH activity,proportion of PI-positive cells and proportion of pyroptotic cells were inhibited;After administration of the NF-κB inhibitor BAY to A549 cells,the effects of B1 on the expression of NF-κB,NLRP3,GSDMD and N-GSDMD were significantly inhibited.Conclusions(1)B1 can inhibit the proliferation,invasion and migration of A549 cells,and inhibit the growth of lung cancer xenograft tumor.(2)B1 plays an antitumor role by inducing caspase-1-dependent cell pyroapoptosis.(3)B1 plays an anti-lung cancer role by regulating NF-κB/NLRP3/ Caspase-1/GSDMD signaling pathway.