DNA methylation downregulated ZDHHC1 suppresses tumor growth by altering cellular metabolism and inducing oxidative/ER stress-mediated apoptosis and pyroptosis
DNA methylation downregulated ZDHHC1 suppresses tumor growth by altering cellular metabolism and inducing oxidative/ER stress-mediated apoptosis and pyroptosis
复制标题
DNA 甲基化下调 ZDHHC1 通过改变细胞代谢和诱导氧化/内质网应激介导的细胞凋亡和焦亡来抑制肿瘤生长
DOI：
10.7150/thno.45631
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Xiang, Tingxiu
中科院分区：
文献类型：
--
作者：
Le, Xin;Mu, Junhao;Xiang, Tingxiu
癌症进展是一个复杂的生物学过程，不仅表现为非计划性增殖，而且还表现为代谢机制的改变。在本文中，我们介绍了一个新的肿瘤抑制基因（TSG），锌指DHHC-Type Containing 1（ZDHHC 1，也称为ZNF 377），在各种癌症中经常由于表观遗传修饰而沉默，通过代谢调节发挥显着的抗肿瘤作用。研究方法：采用定量逆转录PCR（qRT-PCR）、逆转录PCR（RT-PCR）和Western印迹来证明靶向调节物的转录和蛋白水平。采用亚硫酸氢盐基因组测序（BGS）和甲基化特异性PCR（MSP）检测ZDHHC 1启动子甲基化。蛋白质组学分析的同量异位素标签的相对和绝对定量（iTRAQ）和气相色谱-质谱（GC-MS）用于代谢组学分析。通过相应的方法检查细胞功能。裸鼠用于异种移植肿瘤模型。利用间接免疫荧光染色获得靶蛋白的精确定位和表达。使用特定试剂盒检测氧化和ER应激指标。结果：我们发现，由于甲基化，ZDHHC 1在多种肿瘤细胞和标本中的表达经常沉默。ZDHHC 1表达的恢复可以通过刺激细胞凋亡和细胞周期阻滞、抑制转移以及逆转EMT转变和细胞干细胞性来抑制癌细胞的进展。ZDHHC 1的显著抗肿瘤能力在体内也得到了认可。代谢组学和蛋白质组学分析预测了ZDHHC 1以CYGB依赖性方式在葡萄糖代谢途径中以及在磷酸戊糖途径（PPP）中的抑制作用，这一点通过检查改变的关键因素得到了验证。此外，我们揭示了ZDHHC 1致力于增加氧化应激和内质网（ER）应激，以促进细胞凋亡，用于抗癌目的。结论：我们的研究首次表明ZDHHC 1是一种潜在的肿瘤抑制因子，由于启动子甲基化而经常沉默，能够负调节肿瘤细胞的代谢，同时刺激氧化应激和ER应激，通过诱导细胞凋亡和细胞凋亡加速细胞死亡，这可以用于开发新的癌症预防和治疗。
Cancer progression is an intricate biological process profiled by not only unscheduled proliferation, but also altered metabolism mechanisms. In this article, we introduced a novel tumor suppressor gene (TSG), Zinc Finger DHHC-Type Containing 1 (ZDHHC1, also known as ZNF377), frequently silenced due to epigenetic modification among various cancers, which exerts significant anti-tumor effects through metabolic regulation. Methods: Quantitative reversed-transcription PCR (qRT-PCR), reverse transcription PCR (RT-PCR) and Western blot were employed to demonstrate transcriptional and protein levels of targeted regulators. Methylation of ZDHHC1 promoter was detected by bisulfite genomic sequencing (BGS) and methylation specific PCR (MSP). Proteomics were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) and gas chromatography-mass spectrometry (GC-MS) were utilized for metabolomics analysis. Cellular functions were examined via corresponding approaches. Nude mice were used for xenograft tumor models. Indirect immunofluorescence staining was utilized to obtain precise location and expression of target proteins. Oxidative and ER stress indicators were detected using specific kits. Results: We found that ZDHHC1 expression was frequently silenced in multiple tumor cells and specimens due to methylation. Restoration of ZDHHC1 expression can curb cancer cell progression via stimulating apoptosis and cell cycle arrest, repressing metastasis, and reversing EMT transition and cell stemness. ZDHHC1's salient anti-tumor abilities were recognized in vivo as well. Metabolomic and proteomic analyses predicted inhibitory role of ZDHHC1 in glucose metabolism pathways in a CYGB-dependent manner, and in pentose phosphate pathway (PPP), which was validated by examining altered key factors. Moreover, we unraveled that ZDHHC1 dedicates to the increment of oxidative stress and endoplasmic reticulum (ER) stress to promote pyroptosis for anticancer purposes. Conclusion: Our study for the first time indicates ZDHHC1 is a potential tumor-suppressor frequently silenced due to promoter methylation, capable of negatively regulating metabolisms of tumor cells while stimulating oxidative stress and ER stress to expedite cell death through induction of pyroptosis and apoptosis, which can be exploited for development of new cancer prevention and therapies.