血浆中ZNF582甲基化在结直肠癌早期诊断中的可行性研究

目的:探讨血浆锌指蛋白582(zinc finger protein 582,ZNF582)基因甲基化检测用于结直肠癌早期诊断的可行性。方法:选择2021年1月至2022年6月在昆山市中医医院就诊的结直肠癌患者78例(直肠癌组)、结直肠息肉患者62例(结直肠息肉组)为研究对象,另选83例结肠镜检查正常者作为正常对照组。采用甲基化特异性荧光定量PCR检测血浆中ZNF582的甲基化状态,并进行统计学分析。结果:ZNF582在结直肠癌组患者血浆中的甲基化水平显著高于结直肠息肉组和正常对照组(P<0.0001)。ROC曲线分析显示,ZNF582甲基化用于诊断结直肠癌的AUC为0.879(95%CI 0.818~0.925)。ZNF582甲基化对结直肠息肉和结直肠癌的检测灵敏度分别为37.1%、73.1%,对结直肠病变的诊断特异性为90.4%。ZNF582甲基化对不同性别、分期、分化程度以及肿瘤大小的结直肠癌检测灵敏度无显著差异。结论:血浆ZNF582可以作为一种潜在的操作简便的、非侵入性结直肠癌早期诊断替代方案。

ZNF582高甲基化通过调控黏附分子Nectin-3和NRXN3转录促进鼻咽癌转移

背景与目的表观遗传学调节在鼻咽癌(nasopharyngeal carcinoma,NPC)的发生和发展中发挥着重要作用。然而,目前对调控NPC转移的表观遗传学机制尚知之甚少。本研究旨在发现可调节NPC转移的功能基因,并鉴定NPC的治疗靶点。方法采用亚硫酸氢盐处理的焦磷酸测序法分析NPC组织和细胞系中锌指蛋白582(zinc finger protein 582,ZNF582)的甲基化状态。用实时定量逆转录多聚酶链式反应(quantitative reverse transcription-polymerase chain reaction,qRT-PCR)和Western blotting检测ZNF582的表达。通过体外和体内实验探讨NPC中ZNF582的生物学功能。用染色质免疫共沉淀测序(chromatin immunoprecipitation sequencing,ChIP-seq)法筛选ZNF582的靶基因,用ChIP-qPCR和荧光素酶分析进行验证。结果NPC中ZNF582启动子发生了高甲基化,NPC组织和细胞系中ZNF582的mRNA和蛋白表达水平下调。在体外和体内实验中,恢复ZNF582可抑制NPC的迁移、侵袭和转移,而敲减ZNF582可促进NPC的迁移、侵袭和转移。ZNF582直接调节黏附分子Nectin-3和NRXN3的转录和表达。Nectin-3和NRXN3是ZNF582的功能靶分子,恢复或阻断这些基因可逆转ZNF582在NPC转移中的抑癌活性。结论ZNF582通过调控黏附分子Nectin-3和NRXN3的转录和表达在NPC中发挥抑癌基因活性,为NPC的治疗提供了新靶点。

ZNF582及C13orf18基因甲基化与宫颈癌及癌前病变的相关性

目的探讨ZNF582及C13orf18基因甲基化与宫颈癌及癌前病变的相关性。方法通过甲基化敏感性高分辨率熔解曲线法检测宫颈脱落细胞中ZNF582及C13orf18基因甲基化状态,分析ZNF582及C13orf18基因甲基化与宫颈癌和癌前病变的相关性。结果与Normal组相比,甲基化的ZNF582基因(ZNF582m)在CIN3和/或Tumor组中差异均有统计学意义(P<0.05);甲基化的C13orf18基因(C13orf18m)在Tumor组中差异有统计学意义(P<0.05)。CIN3组和Tumor组(CIN3+)患者ZNF582m检测准确率为85.1%,C13orf18m检测CIN3+患者准确率为82.6%。C13orf18m与高危型人乳头瘤病毒(hrHPV)联合检测,比值比(OR)最高(45.6,10.6~196.1,P<0.01)。结论ZNF582及C13orf18基因甲基化状态与CIN3+的宫颈病变程度显著相关,可能是宫颈癌早筛的潜在生物标志物。

ZNF582基因启动子区在结直肠癌组织中的甲基化状态及临床意义

目的探讨ZNF582基因启动子区在结直肠癌组织中的甲基化状态及临床意义。方法采用免疫组化法检测89例结直肠癌组织和癌旁正常组织中ZNF582蛋白表达,甲基化特异性PCR检测ZNF582基因启动子区的甲基化状态,分析ZNF582基因启动子区甲基化与结直肠癌患者临床参数及预后的关系。结果与癌旁正常组织相比,ZNF582蛋白在结直肠癌组织中的表达较低(P<0.05)。结直肠癌组织发生启动子区甲基化比例高于癌旁正常组织(53.9%vs.29.2%)(P<0.05)。ZNF582基因启动子区甲基化与结直肠癌患者TNM分期、淋巴结转移、远处转移和分化程度密切相关(P<0.05)。术后随访3个月~5年,ZNF582基因启动子区甲基化患者的5年生存率低于非甲基化患者(P<0.01)。结论ZNF582基因启动子区在结直肠癌组织中呈现高甲基化状态,与结直肠癌分化和转移密切相关,有助于结直肠癌患者预后的评估。

ZNF582 hypermethylation promotes metastasis of nasopharyngeal carcinoma by regulating the transcription of adhesion molecules Nectin-3 and NRXN3

Background:Epigenetic regulation plays an important role in the development and progression of nasopharyngeal carcinoma(NPC).However,the epigenetic mechanisms underlying NPC metastasis remains poorly understood.We aimed to find functional genes which regulate the metastasis of NPC and identify therapeutic targets for NPC treatment.Methods:Bisulfite pyrosequencing was used to analyze zinc finger protein 582(ZNF582)methylation in NPC tissues and cell lines.Quantitative reverse transcription-polymerase chain reaction(qRT-PCR)and Western blotting were used to determine the expression of ZNF582.In vitro and in vivo experiments were performed to evaluate the biological function of ZNF582 in NPC.ZNF582-targeting genes were identified by chromatin immunoprecipitation sequencing(ChIP-seq)and were confirmed by ChIP-qPCR and luciferase assay.Results:ZNF582 promoter was hypermethylated in NPC,and both the mRNA and protein levels of ZNF582 were down-regulated in NPC tissues and cell lines.The restoration of ZNF582 inhibited NPC migration,invasion,and metastasis,while the knockdown of ZNF582 promoted NPC migration,invasion,and metastasis in vitro and in vivo.ZNF582 directly regulated the transcription and expression of adhesion molecules Nectin-3 and NRXN3.Both Nectin-3 and NRXN3 were identified as functional targets of ZNF582,and the restoration or abrogation of these genes reversed the tumor suppressor effect of ZNF582 in NPC metastasis.Conclusions:ZNF582 acts as a tumor suppressor gene in NPC by regulating the transcription and expression of adhesion molecules Nectin-3 and NRXN3,which may provide novel therapeutic targets for NPC treatment.