19q13 KRAB zinc-finger protein ZNF471 activates MAPK10/JNK3 signaling but is frequently silenced by promoter CpG methylation in esophageal cancer
19q13 KRAB zinc-finger protein ZNF471 activates MAPK10/JNK3 signaling but is frequently silenced by promoter CpG methylation in esophageal cancer
复制标题
食管癌中 19q13 KRAB 锌指蛋白 ZNF471 激活 MAPK10/JNK3 信号传导，但经常被启动子 CpG 甲基化沉默
DOI：
10.7150/thno.35861
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Tao, Qian
中科院分区：
文献类型：
--
作者：
Sun, Ran;Xiang, Tingxiu;Tao, Qian
锌指蛋白（Zinc-finger proteins，ZFPs）是哺乳动物中最大的转录因子家族，参与细胞分化、增殖、凋亡和肿瘤转化等多种生理过程的调控。大约三分之一的ZFP是Krüppel相关盒结构域（KRAB）-ZFP。方法：采用逆转录PCR和甲基化特异性PCR检测ZNF 471基因的表达和甲基化。通过体外和体内实验，研究ZNF 471异位表达对食管鳞状细胞癌（ESCC）细胞的影响及其机制。结果：我们鉴定了19 q13 KRAB-ZFP，ZNF 471，作为ESCC中的甲基化靶点。我们进一步发现ZNF 471在ESCC组织中的表达明显低于癌旁组织，这是由于其启动子CpG甲基化异常所致，并通过甲基化分析和去甲基化剂治疗进一步证实。ZNF 471基因沉默后，ESCC细胞的形态发生明显改变，诱导细胞凋亡和G 0/G1期阻滞，抑制肿瘤细胞集落形成、存活、迁移和侵袭。重要的是，ZNF 471被发现激活MAPK 10/JNK 3和PCDH家族基因的表达，并通过与MAPK 10/JNK 3启动子结合进一步增强MAPK 10信号转导和下游基因表达。结论：我们的研究结果表明，ZNF 471是一个重要的肿瘤抑制因子，在食管癌发生过程中，ZNF 471功能的丧失阻碍了MAPK 10/JNK 3信号转导。
Zinc-finger proteins (ZFPs) are the largest transcription factor family in mammals, involved in the regulation of multiple physiologic processes including cell differentiation, proliferation, apoptosis and neoplastic transformation. Approximately one-third of ZFPs are Krüppel-associated box domain (KRAB)-ZFPs. Methods: ZNF471 expression and methylation were detected by reverse-transcription PCR and methylation-specific PCR. The impact and mechanism of ectopic ZNF471 expression in esophageal squamous cell carcinoma (ESCC) cells was evaluated in vitro and in vivo. Results: We identified a 19q13 KRAB-ZFP, ZNF471, as a methylated target in ESCC. We further found that ZNF471 is significantly downregulated in ESCC tissues compared with adjacent non-cancer tissues, due to its aberrant promoter CpG methylation, and further confirmed by methylation analysis and treatment with demethylation agent. Restoration of ZNF471 expression in silenced ESCC cells significantly altered cell morphology, induced apoptosis and G0/G1 arrest, and inhibited tumor cell colony formation, viability, migration and invasion. Importantly, ZNF471 was found to activate the expression of MAPK10/JNK3 and PCDH family genes, and further enhance MAPK10 signaling and downstream gene expression through binding to the MAPK10/JNK3 promoter. Conclusion: Our results demonstrate that ZNF471 is an important tumor suppressor and loss of ZNF471 functions hampers MAPK10/JNK3 signaling during esophageal carcinogenesis.