Construction Of Oncolytic Adenovirus Carrying GSDME Gene And Its Inhibition Effect On Hepatocellular Carcinoma Cell Growth

At present,the incidence and mortality of liver cancer have increased year by year.In China,the mortality rate of liver cancer in male and female cancer patients ranks second and third place respectively,and it is easy to relapse and metastasize after treatment.Therefore,it is urgent to find a new method for treating liver cancer.Pyroptosis is a new form of cell death,the GSDME gene is a tumor suppressor gene and an important gene for pyroptosis.In order to explore the role of GSDME gene in the targeted therapy of liver cancer,the oncolytic adenovirus GD55-GSDME carrying the GSDME gene was constructed using the previously constructed GP73-regulated oncolytic adenovirus GD55 as a vector and carry through the study on the growth of anti-hepatocellular carcinoma cells by single virus or combined with chemotherapy drug Dox.In this study,the background expression of GSDME gene in hepatocellular carcinoma cells and clinical liver cancer tissues was detected by qRT-PCR,Western blot and immunohistochemistry.The oncolytic adenovirus GD55-GSDME carrying GSDME gene was constructed by genetic engineering.The effects of GD55-GSDME and Dox on the growth of hepatoma cells were detected by MTT,LDH and crystal violet staining.The expressions of GSDME,GSDME-N,caspase 3 and other related proteins were detected by Western blot after treated with GD55-GSDME and Dox.The effect of GD55-GSDME and Dox on apoptosis was detected by Hoechst33342 and flow cytometry.In vivo,the inhibition effect of combined treatment of oncolytic adenovirus GD55-GSDME and Dox was detected by xenotransplantation model.The results showed that when GD55-GSDME and Dox were combined to treat tumor cells,the cells would occur concurrent apoptosis and pyroptosis,and the combined treatment would cause more pyroptosis,which was more effective than any single treatment in inhibiting the growth of liver cancer cells.In summary,we reconstructed a novel oncolytic adenovirus GD55-GSDME regulated by the GP73 promoter and carrying GSDME gene,which can specifically target liver cancer and inhibit the growth of hepatocellular carcinoma cells in vitro and in vivo.It has a good synergistic effect with Dox and less effect on normal cells.It was further found that the oncolytic adenovirus GD55-GSDME combined with Dox can cause apoptosis and pyroptosis of liver cancer cells,which is expected to provide new idea and new technology for the clinical treatment of liver cancer.