Zinc finger and SCAN domain containing 1, ZSCAN1, is a novel stemness-related tumor suppressor and transcriptional repressor in breast cancer targeting TAZ

Jian Chu; Yunzhe Li; Misi He; Hui Zhang; Lingling Yang; Muyao Yang; Jingshu Liu; Chenxi Cui; Liquan Hong; Xingchi Hu; Lei Zhou; Tangya Li; Changchun Li; Huiwen Fan; Guoqin Jiang; Tingyuan Lang

doi:10.3389/fonc.2023.1041688

Zinc finger and SCAN domain containing 1, ZSCAN1, is a novel stemness-related tumor suppressor and transcriptional repressor in breast cancer targeting TAZ

Front Oncol. 2023 Feb 27:13:1041688. doi: 10.3389/fonc.2023.1041688. eCollection 2023.

Authors

Jian Chu¹, Yunzhe Li², Misi He³, Hui Zhang⁴, Lingling Yang⁵, Muyao Yang², Jingshu Liu⁶, Chenxi Cui⁵, Liquan Hong⁷, Xingchi Hu⁸, Lei Zhou^{9

10}, Tangya Li⁸, Changchun Li⁸, Huiwen Fan⁸, Guoqin Jiang¹, Tingyuan Lang^{3

11}

Affiliations

¹ Department of Surgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
² College of Bioengineering, Chongqing University, Chongqing, China.
³ Department of Gynecologic Oncology, Chongqing University Cancer Hospital & Chongqing Cancer Institute & Chongqing Cancer Hospital, Chongqing, China.
⁴ Department of Breast Cancer Center, Chongqing University Cancer Hospital, Chongqing, China.
⁵ School of Medicine, Chongqing University, Chongqing, China.
⁶ Obstetrics and Gynecology Department, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
⁷ Department of Clinical Laboratory, Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.
⁸ Department of General Surgery, Yancheng City No.1 People's Hospital, Yancheng, Jiangsu, China.
⁹ School of Optometry, Department of Applied Biology and Chemical Technology, Research Centre for SHARP Vision (RCSV), The Hong Kong Polytechnic University, Hong Kong, Hong Kong SAR, China.
¹⁰ Centre for Eye and Vision Research (CEVR), 17W Hong Kong Science Park, Hong Kong, Hong Kong SAR, China.
¹¹ Reproductive Medicine Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

Abstract

Introduction: 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.

Methods: 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.

Results and discussion: 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.

Keywords: TAZ; ZSCAN1; breast cancer; stemness; transcriptional repressor; tumor suppressor.

Grants and funding

This study was partially supported by National Natural Science Foundation of China (81873730), Zhejiang medical and health science and technology plan project (2018KY590), Chongqing Science & Technology Commission (CSTB2022NSCQ-MSX1413, cstc2020jxjl130019, cstc2019jscx-msxmX0174), Suzhou Health Planning Commission’s Key Clinical Diagnosis and Treatment Program (LCZX201606), Jiangsu Women and Children Health Key Discipline Program (FXK201758).