| Part Ⅰ Metabolic reprogramming-related genes were screened and ACLY exhibits a significant co-expression relationship with the hub genes in liver cancerObjective To screen metabolic reprogramming-related genes(MRRGs)as candidates,which might be involved in oncogenesis and progression of liver cancer and to identify the co-expression relationship between ACLY and the hub genes in protein-protein interaction(PPI)network in liver cancer.Methods Sequencing data from 50 pairs of tumors and matched normal tissues in TCGA were enrolled in differential expression analysis to screen a list of whole gene expression profiles and a list of differentially expressed genes(DEGs).Then MRRGs were screened through GSEA,GO and KEGG analysis.A protein-protein interaction network was made with the DEGs to identify a co-expression relationship between ACLY and the hub genes in PPI network.Results 26 MRRGs were screened through differential expression analysis and gene enrichment analysis,and ACLY,an MRRG,exhibited a significant co-expression relationship with the hub genes in PPI network.Conclusion 26 MRRGs as candidates might be related to metabolic reprogramming in liver cancer.There is a significant co-expression relationship between ACLY and the hub genes in liver cancer.Part Ⅱ ACLY promotes proliferation and metastasis through endoplasmic reticulum stress in liver cancerObjective To confirm the role of ACLY on proliferation and metastasis of liver cancer and to explore the mechanism.Additionally,to investigate the effect of ACLY inhibition on liver cancer.Methods The expression data of ACLY from TCGA were analyzed.qRT-PCR and western blot were used to detect the expression of ACLY in tumors and matched normal tissues.CCK8 assay and colony formation assay were used to detect the effect of ACLY on proliferation.Wound-healing assay,transwell migration assay and transwell invasion assay were used to establish the effects of ACLY on invasion and metastasis of liver cancer cells.Western blot was used to detect the endoplasmic reticulum stress signaling pathway marker in HepG2 cells stimulated by different concentrations of ACLY inhibitor(ACLYi).The tumor xenograft assay was used to evaluate the effects of ACLYi and sorafenib on the growth inhibition of liver cancer in vivo.Results The expression of ACLY in tumors was significantly higher than that in matched normal tissues in TCGA group.The expression of ACLY in tumors was also significantly higher than that in matched normal tissues in clinical specimen.In vitro,knockdown of ACLY could significantly inhibit proliferation,migration,and invasion of HepG2,overexpression of ACLY could significantly promote proliferation of HCC-LM3.Moreover,cell experiments and tumor xenograft experiment demonstrated that ACLYi induced ERS and initiated apoptosis in liver cancer cells,and that ACLYi synergistically enhanced sorafenib-induced growth inhibition of liver cancer in vivo.Conclusion The expression of ACLY in tumor tissues is higher than that in adjacent tissues,which promotes proliferation and metastasis through ERS in liver cancer.Furthermore,ACLY inhibition can enhance sorafenib-induced growth inhibition of liver cancer in vivo.Part Ⅲ ACLY is an independent risk factor for survival prognosis in patients with liver cancerObjective To establish the correlation between ACLY and clinical pathological indexes and the impact of ACLY on survival of patients with liver cancer.Methods Immunohistochemistry was used to detect the expression of ACLY in tumor tissues.The correlation between ACLY and clinical pathological indexes and the effect of ACLY on prognosis in patients with liver cancer were also analyzed.Additionally,GSE14520 dataset was downloaded from GEO.According to the values of ACLY,221 samples belonging to GPL3921 were divided into high expression group and low expression group.Expression data and clinical data from the 221 patients were enrolled in survival analysis,Cox regression analysis,and clinicopathological analysis.Results The expression level of ACLY was significantly associated with the metastasis of liver cancer(p<0.05).Cox regression analysis showed that ACLY,AFP,TNM staging and tumor size were all independent risk factors for overall survival(p<0.05).Survival analysis showed that overall survival was significantly higher in patients with low ACLY expression than that with high ACLY expression(p<0.05).In GEO group,survival analysis showed that ACLY was significantly correlated with overall survival of patients with liver cancer(p<0.05).Abnormal expression of ACLY was significantly related to TNM staging,CLIP staging,AFP(p<0.05).Cox regression analysis showed that the expression level of ACLY was significantly associated with overall survival and postoperative recurrence-free survival(p<0.05).Conclusion ACLY expression is significantly associated with metastasis and ACLY is an independent prognostic factor for survival in patients with liver cancer.It may be a new target for targeted therapy of liver cancer. |
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