Chu Jian, Li Yunzhe, He Misi, Zhang Hui, Yang Lingling, Yang Muyao, Liu Jingshu, Cui Chenxi, Hong Liquan, Hu Xingchi, Zhou Lei, Li Tangya, Li Changchun, Fan Huiwen, Jiang Guoqin, Lang Tingyuan
Department of Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
College of Bioengineering, Chongqing University, Chongqing, China.
Front Oncol. 2023 Feb 27;13:1041688. doi: 10.3389/fonc.2023.1041688. eCollection 2023.
Cancer stem cells (CSCs) targeted therapy holds the potential for improving cancer management; identification of stemness-related genes in CSCs is necessary for its development.
The Cancer Genome Atlas (TCGA) and the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) datasets were used for survival analysis. ZSCAN1 correlated genes was identified by Spearman correlation analysis. Breast cancer stem-like cells (BCSLCs) were isolated by sorting CD44+CD24- cells from suspension cultured breast cancer (BC) spheroids. The sphere-forming capacity and sphere- and tumor-initiating capacities were determined by sphere formation and limiting dilution assays. The relative gene expression was determined by qRT-PCR, western blot. Lentivirus system was used for gene manipulation. Nuclear run-on assay was employed to examine the levels of nascent mRNAs. DNA pull-down and Chromatin immunoprecipitation (ChIP) assays were used for determining the interaction between protein and target DNA fragments. Luciferase reporter assay was used for evaluating the activity of the promoter.
ZSCAN1 is aberrantly suppressed in BC, and this suppression indicates a bad prognosis. Ectopic expression of ZSCAN1 inhibited the proliferation, clonogenicity, and tumorigenicity of BC cells. ZSCAN1-overexpressing BCSLCs exhibited weakened stemness properties. Normal human mammary epithelial (HMLE) cells with ZSCAN1 depletion exhibited enhanced stemness properties. Mechanistic studies showed that ZSCAN1 directly binds to -951 ~ -925bp region of WWTR1 (encodes TAZ) promoter, inhibits WWTR1 transcription, thereby inhibiting the stemness of BCSCs. Our work thus revealed ZSCAN1 as a novel stemness-related tumor suppressor and transcriptional repressor in BC.
癌症干细胞（CSCs）靶向治疗具有改善癌症治疗效果的潜力；鉴定CSCs中与干性相关的基因对其发展至关重要。
使用癌症基因组图谱（TCGA）和国际乳腺癌分子分类联盟（METABRIC）数据集进行生存分析。通过Spearman相关分析鉴定与ZSCAN1相关的基因。从悬浮培养的乳腺癌（BC）球体中筛选CD44 + CD24-细胞来分离乳腺癌干细胞样细胞（BCSLCs）。通过成球实验和极限稀释实验确定成球能力以及球体形成和肿瘤起始能力。通过qRT-PCR、蛋白质免疫印迹法测定相对基因表达。利用慢病毒系统进行基因操作。采用核转录分析检测新生mRNA水平。DNA下拉实验和染色质免疫沉淀（ChIP）实验用于确定蛋白质与靶DNA片段之间的相互作用。荧光素酶报告基因实验用于评估启动子活性。
ZSCAN1在BC中异常抑制，这种抑制表明预后不良。ZSCAN1的异位表达抑制了BC细胞的增殖、克隆形成能力和致瘤性。过表达ZSCAN1的BCSLCs表现出减弱的干性特征。ZSCAN1缺失的正常人乳腺上皮（HMLE）细胞表现出增强的干性特征。机制研究表明，ZSCAN1直接结合WWTR1（编码TAZ）启动子的-951 ~ -925bp区域，抑制WWTR1转录，从而抑制BCSCs的干性。因此，我们的研究揭示了ZSCAN1是BC中一种新型的与干性相关的肿瘤抑制因子和转录抑制因子。
