DNA methylation of RUNX3 promotes the progression of gallbladder cancer through repressing SLC7A11-mediated ferroptosis
DNA methylation of RUNX3 promotes the progression of gallbladder cancer through repressing SLC7A11-mediated ferroptosis
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DOI：
10.1016/j.cellsig.2023.110710
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发表时间：
2023-05-10
影响因子：
4.8
通讯作者：
Gong，Wei
中科院分区：
文献类型：
--
作者：
Cai，Chen;Zhu，Yidi;Gong，Wei
胆囊癌（GBC）是一种罕见但高度侵袭性的癌症，预后不良。Runt-related transcription factor 3（RUNX 3）是Runt-domain家族的成员之一，其启动子甲基化在多种恶性肿瘤中广泛存在。然而，RUNX 3在GBC中的生物学功能和潜在机制仍不清楚。本研究采用硫酸氢盐测序PCR（BSP）、Western blot和qPCR检测了RUNX 3在胆囊癌组织和细胞中的表达水平和DNA甲基化水平。通过双荧光素酶报告基因检测和ChIP检测验证了RUNX 3和生长抑制因子1（Inhibitor of growth 1，ING 1）之间的转录关系。通过一系列的功能获得和功能丧失试验，检测RUNX 3在体外和体内的功能和调控关系。DNA甲基转移酶1（DNMT 1）介导的甲基化导致RUNX 3在GBC细胞和组织中异常下调，RUNX 3的下调与GBC患者的不良预后相关。功能实验表明，RUNX 3可以诱导GBC细胞在体外和体内的铁凋亡。机制上，RUNX 3通过激活ING 1转录诱导铁凋亡，从而以p53依赖性方式抑制SLC 7A 11。总之，RUNX 3的下调是由DNA甲基化介导的，DNA甲基化通过减弱SLC 7A 11介导的铁凋亡促进胆囊癌的发病。这项研究为RUNX 3在GBC细胞铁凋亡中的作用提供了新的见解，这可能有助于开发GBC的潜在治疗靶点。
Gallbladder cancer (GBC) is a type of rare but highly aggressive cancer with a dismal prognosis. Runt-related transcription factor 3 (RUNX3), a member of the runt-domain family, and its promoter methylation have been widely observed in a variety of human malignancies. However, the biological function and underlying mechanism of RUNX3 in GBC remain elusive. In this study, bisulfate sequencing PCR (BSP), Western blot, and qPCR were applied to identify the expression level and DNA methylation level of RUNX3 in GBC tissues and cells. The transcriptional relationship between RUNX3 and Inhibitor of growth 1 (ING1) was validated by dual-luciferase reporter assay and ChIP assay. A series of gain-of-function and loss-of-function assays were performed to detect the function and the regulatory relationship of RUNX3 in vitro and in vivo. RUNX3 was aberrantly downregulated in GBC cells and tissues caused by DNA Methyltransferase 1 (DNMT1)-mediated methylation, and downregulation of RUNX3 is associated with poor prognosis of GBC patients. Functional experiments reveal that RUNX3 can induce ferroptosis of GBC cells in vitro and in vivo. Mechanistically, RUNX3 induces ferroptosis by activating ING1 transcription, thereby repressing SLC7A11 in a p53-dependent manner. In conclusion, the downregulation of RUNX3 is mediated by DNA methylation, which promotes the pathogenesis of gallbladder cancer through attenuating SLC7A11-mediated ferroptosis. This study gives novel insights into the role of RUNX3 in the ferroptosis of GBC cells, which may contribute to developing potential treatment targets for GBC.