Baculoviral IAP repeat containing 5 influences the proliferation of adrenocortical carcinoma by acti-vating the mammalian target of rapamycin pathway
Objective To investigate the effects of baculoviral IAP repeat containing 5(BIRC5)on autophagy and proliferation in adrenocortical carcinoma cells and its underlying mechanisms.Methods An analysis of differential genes in 77 adrenocortical carcinoma tissues and 128 normal adrenal tissues from the TCGA and GTEx databases identified 5 upregulated and 25 downregulated genes at the intersection of differential and autophagy-related genes.Survival analysis was performed on the enriched genes,followed by single-gene analysis of the gene with the most significant survival difference.HUB gene extraction was conducted using Cytoscape and STRING,and gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)analyses were carried out.Human adrenocortical carcinoma cell lines SW-13 and NCL-H295 were used as study models,transfected with either empty plasmids or BIRC5 overexpression plas-mids.Two days after transfection,a colony formation assay was used to assess cell proliferation.Western blotting analysis was employed to explore the relationship between BIRC5,autophagy,and p53.An inde-pendent sample t-test was used for comparisons between the two experimental groups.Results BIRC5 showed the most significant survival difference(P＜0.01).The number of cell colonies in the BIRC5 over-expression group was higher than in the empty plasmid group(SW-13:832.000±29.933 vs.575.333±19.619,t=10.140,P＜0.05;NCL-H295:696.667±30.126 vs.519.667±15.923,t=7.346,P＜0.05).In the group treated with the p53 activator flubendazole along with BIRC5 overexpression,the num-ber of colonies was lower than in the BIRC5 overexpression group(SW-13:560.667±26.247 vs.675.667±13.021,t=5.551,P＜0.05;NCL-H295:521.667±20.886 vs.747.000±62.759,t=4.818,P＜0.05).The expression of p53 protein in the overexpressed BIRC5 group was significantly lower than that in the no-loaded plasmid group(SW-13:0.610±0.093 vs.0.855±0.031,t=3.556,P＜0.05;NCL-H295:0.596±0.102 vs.0.859±0.042,t=3.390,P＜0.05),increased phosphorylated mammalian target of rapamycin(p-mTOR)protein expression(SW-13:0.867±0.024 vs.0.612±0.098,t=3.575,P＜0.05;NCL-H295:0.947±0.142 vs.0.450±0.109,t=3.911,P＜0.05),and decreased LC3B protein expression(SW-13:0.657±0.166 vs.1.136±0.110,t=3.408,P＜0.05;NCL-H295:0.391±0.040 vs.1.116±0.089,t=10.540,P＜0.05).The expression of p53 protein in the over-expression BIRC5 group was significantly higher than that in the over-expression BIRC5 group(SW-13:1.074±0.037 vs.0.610±0.093,t=6.582,P＜0.05;NCL-H295:1.087±0.108 vs.0.596±0.102,t=4.681,P＜0.05),p-mTOR protein expression decreased(SW-13:0.520±0.091 vs.0.872±0.081,t=4.088,P＜0.05;NCL-H295:0.570±0.133 vs.1.062±0.019,t=5.159,P＜0.05),and LC3B protein expression increased(SW-13:1.194±0.085 vs.0.514±0.082,t=8.143,P＜0.05;NCL-H295:1.345±0.215 vs.0.483±0.028,t=5.617,P＜0.05).Conclusion BIRC5 inhibits auto-phagy and promotes the proliferation of adrenocortical carcinoma via the p53/mTOR axis.
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