The Effect Of GSDMD On The Biological Function Of Hepatocellular Carcinoma Cells And Its Mechanism

Objective: Liver cancer is associated with poor prognosis due to inefficient early diagnosis and rapid progression,and traditional treatment relies on surgical resection and radiotherapy,but most patients are at risk of recurrence and distant metastasis after treatment.Currently,an increasing number of studies have shown that tumor progression is associated with inflammatory changes in the immune microenvironment.Pyroptosis,a form of programmed cell death distinct from apoptosis,is characterized by membrane pore formation and the maturation and release of pro-inflammatory cell contents containing IL-1β and IL-18.As an important component of immune regulation,it occurs in varying degrees by modulating antitumor immune potency to influence tumor progression.In pyroptosis,Gasdermin D(GSDMD)is a key effector protein that regulates the onset of pyroptosis.Activation of caspase-1 cleaves GSDMD to produce the GSDMD-N terminus,which binds to the cell membrane to create membrane pores and promote cell expansion and rupture to release large amounts of cellular contents and induce the activation of inflammatory factors such as IL-1β and IL-18.and other inflammatory factors,and finally cause a severe cellular inflammatory response.However,current studies have focused on the effects of GSDMD-N-mediated pyroptosis on tumor microenvironment(TME)remodeling and tumor progression,while neglecting the role of GSDMD on tumor proliferation and migration.In addition,mitochondria serve as the center of metabolic activity,providing nutrients and energy for tumor development,and it is unknown whether GSDMD can regulate tumor progression by affecting mitochondrial function.Therefore,we aimed to investigate the effect of GSDMD itself on tumor progression and to experimentally analyze in which way GSDMD is involved in regulating hepatocellular carcinoma progression,with the aim of providing a reference for tumor-targeted therapy against GSDMD.Methods: The pan-cancer transcriptome data were cleaned and normalized to analyze the differential expression of GSDMD in pan-cancer and its prognostic correlation,and to compare the association and statistical significance between the expression levels of GSDMD in different tumors and genomic instability,DNA repair,cancer stemness,RNA epigenetic modifications,immune infiltration and drug sensitivity.The prognosis-related genes of hepatocellular carcinoma were intersected with mitochondrial function-related genes and their correlation with GSDMD was analyzed.The gene with the most correlation with GSDMD was selected for follow-up experiments,and the two genes were knocked down separately to observe their effects on proliferation,migration and invasion of hepatocellular carcinoma,and the relationship between GSDMD and RECQL4 was verified by Western Blotting.Results: Bioinformatics analysis suggested that GSDMD was statistically significantly differentially expressed in hepatocellular carcinoma(HCC),breast cancer(BRCA),glioblastoma multiforme(GBM),gastric cancer(STAD)and other tumors(P <0.05),and high expression of GSDMD was associated with poor prognosis in hepatocellular carcinoma and other tumors,which may be due to the fact that high expression of GSDMD affects tumor This may be due to the fact that high expression of GSDMD is involved in tumor progression by affecting tumor mutation burden(TMB),purity(a marker of chromosomal instability),DNA mismatch repair,epigenetics(m1A,m5 C,m6A),immune infiltration,chemokines and other factors.GSDMD is highly expressed in hepatocellular carcinoma cells such as Huh7 compared to normal hepatocytes and GSDMD knockdown in hepatocellular carcinoma cells inhibits proliferation,migration and invasion of hepatocellular carcinoma cells.RECQL4 is a mitochondrial-associated gene whose expression maintains the functional integrity of mitochondria and promotes the proliferation,migration and invasion of hepatocellular carcinoma,and GSDMD may coregulate hepatocellular carcinoma progression with RECQL4.Conclusion: GSDMD and RECQL4 combine to promote the proliferation,migration and invasion of hepatocellular carcinoma cells.