Yoon Jung Hwan, Kim Jeong-Kyu, Eun Jung Woo, Ashktorab Hassan, Smoot Duane T, Nam Suk Woo, Park Won Sang
Department of Pathology, Functional RNomics Research Center, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 06591, South Korea.
Department of Life Science, Chung-Ang University, Seoul, South Korea.
Cell Commun Signal. 2025 Jan 20;23(1):35. doi: 10.1186/s12964-025-02030-4.
Gastric cancer remains a significant global health challenge, characterized by poor prognosis and high mortality rates. Mitotic integrity and genomic stability are crucial in maintaining cellular homeostasis and preventing tumorigenesis. The transcription factor NKX6.3 has emerged as a potential regulator of these processes in gastric epithelial cells, prompting an investigation into its role in gastric cancer development.
We employed a combination of in vitro and in vivo techniques to elucidate the impact of NKX6.3 depletion on mitotic dynamics and genomic stability in gastric epithelial cells. Quantitative real-time PCR and Western blot analyses were conducted to assess the expression of mitosis-related genes and proteins. Flow cytometry was utilized to evaluate cell cycle distribution, while immunofluorescence microscopy enabled the visualization of mitotic abnormalities. Statistical analyses, including Student's t-test and ANOVA, were performed to determine the significance of our findings.
Our results demonstrate that NKX6.3 depletion leads to significant mitotic defects, characterized by increased chromosome misalignment and lagging chromosomes during anaphase. These abnormalities corresponded with elevated levels of genomic instability markers, indicating compromised genomic integrity. Furthermore, the loss of NKX6.3 resulted in altered expression of key regulatory proteins involved in mitosis and DNA repair pathways, suggesting a mechanistic link between NKX6.3 and the maintenance of genomic stability in gastric epithelial cells. Depletion of NKX6.3 resulted in accelerated cell cycle progression and the formation of abnormal mitotic figures, leading to genomic instability characterized by increased DNA content and structural abnormalities. In both in vitro and xenograft models, the depletion of NKX6.3 significantly upregulated AurkA and TPX2, which correlated with gains in DNA copy number. An inverse relationship was observed between NKX6.3 expression and the levels of AurkA and TPX2 in human gastric cancer tissues.
This study highlights the essential role of NKX6.3 in regulating mitotic integrity and genomic stability in gastric carcinogenesis. The findings suggest that targeting NKX6.3 may offer a novel therapeutic strategy for improving treatment outcomes in gastric cancer by restoring mitotic fidelity and genomic stability.
This study was not registered.
胃癌仍然是一项重大的全球健康挑战，其特点是预后不良和死亡率高。有丝分裂完整性和基因组稳定性对于维持细胞内稳态和预防肿瘤发生至关重要。转录因子NKX6.3已成为胃上皮细胞中这些过程的潜在调节因子，促使人们对其在胃癌发展中的作用进行研究。
我们采用体外和体内技术相结合的方法，以阐明NKX6.3缺失对胃上皮细胞有丝分裂动力学和基因组稳定性的影响。进行定量实时PCR和蛋白质印迹分析以评估有丝分裂相关基因和蛋白质的表达。利用流式细胞术评估细胞周期分布，而免疫荧光显微镜能够观察到有丝分裂异常。进行包括学生t检验和方差分析在内的统计分析，以确定我们研究结果的显著性。
我们的结果表明，NKX6.3缺失导致显著的有丝分裂缺陷，其特征是后期染色体错配和滞后染色体增加。这些异常与基因组不稳定标志物水平升高相对应，表明基因组完整性受损。此外，NKX6.3的缺失导致参与有丝分裂和DNA修复途径的关键调节蛋白表达改变，提示NKX6.3与胃上皮细胞基因组稳定性维持之间存在机制联系。NKX6.3的缺失导致细胞周期进程加速和异常有丝分裂图像的形成，导致以DNA含量增加和结构异常为特征的基因组不稳定。在体外和异种移植模型中，NKX6.3的缺失均显著上调AurkA和TPX2，这与DNA拷贝数增加相关。在人胃癌组织中观察到NKX6.3表达与AurkA和TPX2水平之间呈负相关。
本研究强调了NKX6.3在胃癌发生过程中调节有丝分裂完整性和基因组稳定性的重要作用。研究结果表明，靶向NKX6.3可能通过恢复有丝分裂保真度和基因组稳定性为改善胃癌治疗结果提供一种新的治疗策略。
本研究未注册。
