Li Xiaowei, Li Xubin, Wang Jing, Xue Mei, Zhang Mengqiao, Shuai Junfang, Zhang Jie, Ding Danyang, Wang Ye, Hou Shiyang, Chi Xiaoqian, Sun Haiying, Gao Qiang, Kang Chunbo
Department of Surgery, Center of Gastrointestinal Rehabilitation, Beijing Rehabilitation Hospital, Capital Medical University, Beijing 100144, P.R. China.
Department of Gastroenterology and Hepatology, Center of Gastrointestinal Rehabilitation, Beijing Rehabilitation Hospital, Capital Medical University, Beijing 100144, P.R. China.
Oncol Rep. 2025 Oct;54(4). doi: 10.3892/or.2025.8949. Epub 2025 Jul 19.
Colorectal cancer (CRC) is the third most common malignant tumor and the second leading cause of cancer‑related deaths worldwide. Identifying driver genes in CRC development may provide clinical benefits for patients. Zinc finger protein 695 (ZNF695) is a nuclear protein with transcriptional regulatory activity, which has been implicated in tumor progression; however, the role of ZNF695 in CRC is unclear. The clinical relevance of ZNF695 and NIMA‑related kinase 2 (NEK2) in patients with CRC was analyzed based on The Cancer Genome Atlas database. Knockdown was performed by transfecting the cells with small interfering RNAs, whereas overexpression was induced by infecting the cells with a lentivirus. In addition, cell Counting Kit‑8, colony formation, cell cycle and apoptosis assays were carried out to assess the role of ZNF695 and NEK2 in CRC. Chromatin immunoprecipitation‑quantitative PCR (qPCR) and dual luciferase reporter assays were used to examine the transcriptional regulation of ZNF695 on the NEK2 gene. Reverse transcription‑qPCR and western blotting were applied to assess mRNA and protein abundance, respectively. The present study aimed to investigate the clinical relevance, contribution and downstream effects of ZNF695 in CRC. The results revealed that ZNF695 was upregulated in CRC tissues compared with that in non‑cancer tissues. CRC cells also expressed higher ZNF695 expression than normal cells. , knockdown of ZNF695 suppressed the proliferation of HCT‑8 cells; conversely, overexpression of ZNF695 promoted the malignancy of HT‑29 cells. Moreover, ZNF695 accelerated cell cycle progression and inhibited apoptosis in CRC cells. Mechanistically, it was revealed that ZNF695 upregulated the expression of NEK2 at both the mRNA and protein levels. Luciferase reporter assay demonstrated that ZNF695 enhanced the transcriptional activity of the NEK2 promoter. Furthermore, knockdown of NEK2 reversed the oncogenic function of ZNF695. Additionally, ZNF695 activated the PI3K/Akt/mTOR signaling pathway in CRC cells. Inhibition of this pathway with rapamycin resulted in higher cytotoxicity to CRC cells with ZNF695 overexpression, suggesting that elevated ZNF695 levels may increase the sensitivity of CRC cells to rapamycin. In summary, the current study identified ZNF695 as a tumor‑promoting protein in CRC through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways. Targeting the NEK2 and PI3K/Akt/mTOR signaling pathways may therefore be a promising strategy for the treatment of patients with CRC and high ZNF695 expression.
结直肠癌（CRC）是全球第三大常见恶性肿瘤，也是癌症相关死亡的第二大主要原因。确定CRC发生过程中的驱动基因可能会给患者带来临床益处。锌指蛋白695（ZNF695）是一种具有转录调节活性的核蛋白，与肿瘤进展有关；然而，ZNF695在CRC中的作用尚不清楚。基于癌症基因组图谱数据库分析了ZNF695和NIMA相关激酶2（NEK2）在CRC患者中的临床相关性。通过用小干扰RNA转染细胞进行敲低，而通过用慢病毒感染细胞诱导过表达。此外，进行细胞计数试剂盒8、集落形成、细胞周期和凋亡检测以评估ZNF695和NEK2在CRC中的作用。采用染色质免疫沉淀定量PCR（qPCR）和双荧光素酶报告基因检测来研究ZNF695对NEK2基因的转录调控。分别应用逆转录qPCR和蛋白质印迹法评估mRNA和蛋白质丰度。本研究旨在探讨ZNF695在CRC中的临床相关性、作用及下游效应。结果显示，与非癌组织相比，ZNF695在CRC组织中上调。CRC细胞中ZNF695的表达也高于正常细胞。敲低ZNF695可抑制HCT-8细胞的增殖；相反，ZNF695的过表达促进HT-29细胞的恶性程度。此外，ZNF695加速CRC细胞的细胞周期进程并抑制其凋亡。机制上，发现ZNF695在mRNA和蛋白质水平上均上调NEK2的表达。荧光素酶报告基因检测表明ZNF695增强了NEK2启动子的转录活性。此外，敲低NEK2可逆转ZNF695的致癌功能。另外，ZNF695激活CRC细胞中的PI3K/Akt/mTOR信号通路。用雷帕霉素抑制该通路会导致对过表达ZNF695的CRC细胞具有更高的细胞毒性，这表明ZNF695水平升高可能会增加CRC细胞对雷帕霉素的敏感性。总之，本研究通过激活NEK2和PI3K/Akt/mTOR信号通路确定ZNF695为CRC中的一种促肿瘤蛋白。因此，靶向NEK2和PI3K/Akt/mTOR信号通路可能是治疗ZNF695高表达的CRC患者的一种有前景的策略。
