目的:常染色体显性遗传性多囊肾病(ADPKD)是终末期肾病的最常见原因。已经显示,在ADPKD患者的肾囊性组织中,酰基辅酶A硫酯酶13(ACOT13)水平降低。然而,ACOT13在ADPKD中的作用在很大程度上仍然难以捉摸。
方法:从GEO数据库获取GSE7869数据集中的数据,以确定正常肾皮质组织和肾囊组织之间的ACOT13水平。接下来,通过基因集富集分析(GSEA)探讨了ACOT13的潜在功能。此外,通过RT-qPCR验证ADPKD细胞(WT9-12)中的ACOT13水平。使用EdU染色和流式细胞术测定评价ACOT13对WT9-12细胞生长的影响。
结果:与正常组相比,肾囊组织和WT9-12细胞中ACOT13mRNA水平明显降低。同时,GSEA成果显示,与ACOT13低表达组比拟,PI3K-Akt和MAPK信号通路被灭活,ACOT13高表达组PPARα信号通路和脂肪酸代谢均被激活。此外,过表达ACOT13显著降低WT9-12细胞增殖并触发细胞周期阻滞。此外,ACOT13过表达显著触发细胞凋亡,增加裂解的caspase3蛋白水平,WT9-12细胞中ATP的产生减少和线粒体膜电位的诱导损失,提示ACOT13过表达可引发WT9-12细胞线粒体相关凋亡。
结论:总的来说,我们的结果表明,过表达ACOT13可以抑制WT9-12细胞的增殖并触发线粒体介导的细胞凋亡,提示ACOT13可能在ADPKD中发挥保护作用。
OBJECTIVE: Autosomal dominant polycystic kidney disease (ADPKD) is the most common cause of end-stage kidney disease. It has been shown that Acyl-CoA thioesterase 13 (ACOT13) level was reduced in renal cystic tissues from ADPKD patients. However, the role of ACOT13 in ADPKD remains largely elusive.
METHODS: The data in the GSE7869 dataset were acquired from the GEO database to determine ACOT13 level between normal renal cortical tissues and renal cystic tissues. Next, the potential functions of ACOT13 were explored by gene set enrichment analysis (GSEA). Furthermore, ACOT13 level in ADPKD cells (WT9-12) was verified by RT-qPCR. The effects of ACOT13 on WT9-12 cell growth were evaluated using the EdU staining and flow cytometry assays.
RESULTS: Compared to normal group, ACOT13 mRNA level was obviously reduced in renal cystic tissues and WT9-12 cells. Meanwhile, GSEA results showed that compared to the low ACOT13 expression group, PI3K-Akt and MAPK signaling pathways were inactivated, and PPAR signaling pathway and fatty acid metabolism were activated in high ACOT13 expression group. Furthermore, overexpression of ACOT13 notably reduced WT9-12 cell proliferation and triggered cell cycle arrest. Moreover, ACOT13 overexpression remarkably triggered apoptosis, increased cleaved caspase 3 protein level, reduced ATP production and induced loss of mitochondrial membrane potential in WT9-12 cells, suggesting that ACOT13 overexpression could trigger mitochondrial-related apoptosis in WT9-12 cells.
CONCLUSIONS: Collectively, our results showed that overexpression of ACOT13 could suppress WT9-12 cell proliferation and trigger mitochondrial-mediated cell apoptosis, suggesting that ACOT13 may exert a protective role in ADPKD.