Liu Jianfeng, Li Yuanyuan, Liao Bingling, Xu Qihua, Zhou Ying, Zhang Huijun
Department of Gastroenterology, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, No. 358, Datong Road, Pudong New Area, Shanghai, China.
Cytotechnology. 2025 Jun;77(3):111. doi: 10.1007/s10616-025-00765-z. Epub 2025 May 30.
Colorectal cancer (CRC) is a common malignancy of the digestive tract and is the second leading cause of cancer-related death worldwide. Keratin 23 (KRT23), a member of the keratin family, is implicated in the development of various cancers. Hence, this study aimed to clarify the molecular mechanism of KRT23 in the progression of CRC. Quantitative real time polymerase chain reaction (qRT-PCR) and western blot were applied to detected RNA and protein levels. Cell migration, invasion, and apoptosis were measured by wound healing, transwell assay, and flow cytometry, respectively. Glycolysis was reflected by the detection of ATP, lactate production, and glucose consumption. What's more, the binding site of zinc finger protein 384 (ZNF384) and KRT23 was predicted using the relevant website and verified by chromatin immunoprecipitation (CHIP) and dual-luciferase reporter assay. Meanwhile, the related proteins were detected by immunohistochemistry (IHC). A mouse xenograft model was established for in vivo analysis and further verified the role of ZNF384 and KRT23 in CRC. KRT23 was highly expressed in CRC tissues and cells. Functionally, silencing KRT23 inhibited CRC migration and invasion, promoted apoptosis, and impeded epithelial-mesenchymal transition (EMT) and glycolysis process by the TGF-β/Smad signaling pathway. In terms of mechanism, ZNF384 bound to the KRT23 promoter, and the inhibition caused by ZNF384 interference was reversed with KRT23. In vivo, knocking down ZNF384 inhibited tumor growth. In summary, ZNF384 could promote the malignant progression of CRC by regulating the KRT23-mediated TGF-β/Smad signaling pathway.
The online version contains supplementary material available at 10.1007/s10616-025-00765-z.
结直肠癌（CRC）是消化道常见的恶性肿瘤，是全球癌症相关死亡的第二大主要原因。角蛋白23（KRT23）是角蛋白家族的成员，与多种癌症的发生发展有关。因此，本研究旨在阐明KRT23在CRC进展中的分子机制。应用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测RNA和蛋白质水平。分别通过伤口愈合实验、Transwell实验和流式细胞术检测细胞迁移、侵袭和凋亡情况。通过检测ATP、乳酸生成和葡萄糖消耗来反映糖酵解情况。此外，利用相关网站预测锌指蛋白384（ZNF384）与KRT23的结合位点，并通过染色质免疫沉淀（CHIP）和双荧光素酶报告基因实验进行验证。同时，通过免疫组织化学（IHC）检测相关蛋白。建立小鼠异种移植模型进行体内分析，进一步验证ZNF384和KRT23在CRC中的作用。KRT23在CRC组织和细胞中高表达。在功能上，沉默KRT23可抑制CRC迁移和侵袭，促进凋亡，并通过TGF-β/Smad信号通路阻碍上皮-间质转化（EMT）和糖酵解过程。在机制方面，ZNF384与KRT23启动子结合，ZNF384干扰引起的抑制作用可被KRT23逆转。在体内，敲低ZNF384可抑制肿瘤生长。综上所述，ZNF384可通过调节KRT23介导的TGF-β/Smad信号通路促进CRC的恶性进展。
在线版本包含可在10.1007/s10616-025-00765-z获取的补充材料。
