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.

中文翻译：

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LncRNA ZNF503-AS1 通过转录因子 GATA6 上调 Ca 2+ 浓度作为膀胱癌的肿瘤抑制因子

目的

Ca ²⁺稳态在调节癌症发展过程中的增殖和凋亡中起着关键作用。本研究旨在检查 ZNF503 反义 RNA 1 (ZNF503-AS1) 在膀胱癌中的潜在肿瘤抑制作用，这可能与 Ca ²⁺稳态的调节有关。

方法

使用微阵列分析鉴定与膀胱癌相关的差异表达的长链非编码 RNA (lncRNA)，然后验证它们结合的转录因子。ZNF503-AS1、GATA6 和 SLC8A1 之间的关系使用双荧光素酶报告基因、RIP 和 ChIP 分析进行评估。调节 ZNF503-AS1、GATA6 和 SLC8A1 的表达水平以检查它们对膀胱癌细胞中的致瘤潜力、细胞内 Ca ²⁺浓度和 Ca ²⁺ -ATPase 活性的影响。在裸鼠中验证了体内致瘤能力。

结果

基于微阵列的 GEO GSE61615 数据集的表达谱分析显示 ZNF503-AS1 的表达在膀胱癌中降低。随后，我们发现ZNF503-AS1可以与转录因子GATA6结合，上调SLC8A1的表达。发现 ZNF503-AS1 和 SLC8A1 在原发性膀胱癌组织和细胞中均下调。ZNF503-AS1或SLC8A1的外源性过表达减弱了膀胱癌细胞的增殖、侵袭和迁移，但促进了它们的凋亡，伴随着Ca ²⁺ -ATP酶活性降低和细胞内Ca ²⁺浓度增加。额外的体内实验验证了 ZNF503-AS1 过表达对裸鼠膀胱癌细胞致瘤能力的抑制作用。

结论

ZNF503-AS1可以招募转录因子GATA6上调SLC8A1的表达，从而增加细胞内Ca ²⁺浓度，抑制增殖、侵袭和迁移，增强膀胱癌细胞的凋亡。