LncRNA ZNF503-AS1 acts as a tumor suppressor in bladder cancer by up-regulating Ca2+ concentration via transcription factor GATA6

Haiqing He; Shuiqing Wu; Kai Ai; Ran Xu; Zhaohui Zhong; Yinhuai Wang; Lei Zhang; Xiaokun Zhao; Xuan Zhu

doi:10.1007/s13402-020-00563-z

LncRNA ZNF503-AS1 acts as a tumor suppressor in bladder cancer by up-regulating Ca²⁺ concentration via transcription factor GATA6

Cell Oncol (Dordr). 2021 Feb;44(1):219-233. doi: 10.1007/s13402-020-00563-z. Epub 2020 Oct 1.

Authors

Haiqing He¹, Shuiqing Wu¹, Kai Ai¹, Ran Xu¹, Zhaohui Zhong¹, Yinhuai Wang¹, Lei Zhang¹, Xiaokun Zhao², Xuan Zhu³

Affiliations

¹ Department of Urology, The Second Xiangya Hospital, Central South University, No.139 Renmin Road, Changsha, 410011, Hunan Province, People's Republic of China.
² Department of Urology, The Second Xiangya Hospital, Central South University, No.139 Renmin Road, Changsha, 410011, Hunan Province, People's Republic of China. xiaokunzhao@csu.edu.cn.
³ Department of Urology, The Second Xiangya Hospital, Central South University, No.139 Renmin Road, Changsha, 410011, Hunan Province, People's Republic of China. zhuxuan@csu.edu.cn.

PMID: 33001357
DOI: 10.1007/s13402-020-00563-z

Abstract

Purpose: Ca²⁺ homeostasis plays a pivotal role in regulating proliferation and apoptosis during cancer development. This study intended to examine the potential tumor-suppressing role of ZNF503 antisense RNA 1 (ZNF503-AS1) in bladder cancer, which may be implicated in the regulation of Ca²⁺ homeostasis.

Methods: Differentially expressed long non-coding RNAs (lncRNAs) related to bladder cancer were identified using microarray analysis, followed by the verification of transcription factors to which they bind. The relationship between ZNF503-AS1, GATA6 and SLC8A1 was assessed using dual luciferase reporter, RIP and ChIP assays. The expression levels of ZNF503-AS1, GATA6 and SLC8A1 were modulated to examine their effects on the tumorigenic potential, intracellular Ca²⁺ concentration and Ca²⁺-ATPase activity in bladder cancer cells. The in vivo tumorigenic ability was validated in nude mice.

Results: Microarray-based expression profile analysis of the GEO GSE61615 dataset revealed that the expression of ZNF503-AS1 was decreased in bladder cancer. Subsequently, we found that ZNF503-AS1 can bind to the transcription factor GATA6 to up-regulate the expression of SLC8A1. ZNF503-AS1 and SLC8A1 were found to be down-regulated in both primary bladder cancer tissues and cells. Exogenous overexpression of ZNF503-AS1 or SLC8A1 attenuated bladder cancer cell proliferation, invasion and migration, but promoted their apoptosis, accompanied by decreased Ca²⁺-ATPase activities and increased intracellular Ca²⁺ concentrations. Additional in vivo experiments validated the inhibitory effect of ZNF503-AS1 overexpression on the tumorigenic capacity of bladder cancer cells in nude mice.

Conclusion: ZNF503-AS1 can recruit transcription factor GATA6 to up-regulate SLC8A1 expression, thereby increasing the intracellular Ca²⁺ concentration and repressing the proliferation, invasion and migration, and enhancing the apoptosis of bladder cancer cells.

Keywords: Bladder cancer; Ca2+ concentration; GATA6; Invasion and migration; LncRNA; Proliferation; SLC8A1; Transcription factor; ZNF503-AS1.

MeSH terms

Animals
Apoptosis / genetics
Calcium / metabolism*
Cell Line, Tumor
Cell Movement / genetics
Cell Proliferation / genetics
Down-Regulation / genetics
GATA6 Transcription Factor / metabolism*
Gene Expression Regulation, Neoplastic
Genes, Tumor Suppressor*
Humans
Male
Mice
Mice, Nude
Models, Biological
Neoplasm Invasiveness
RNA, Long Noncoding / genetics
RNA, Long Noncoding / metabolism*
Sodium-Calcium Exchanger / metabolism
Up-Regulation / genetics*
Urinary Bladder Neoplasms / genetics*
Urinary Bladder Neoplasms / pathology

Substances

GATA6 Transcription Factor
RNA, Long Noncoding
Sodium-Calcium Exchanger
long non-coding RNA ZNF503-AS1, human
sodium-calcium exchanger 1
Calcium