Zhang Chunlei, Yi Shiming, Cao Xiansheng, Wang Jiafeng
Department of Colorectal and Anus Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Department of Hepatobiliary Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Iran J Allergy Asthma Immunol. 2025 Jun 26;24(4):481-497.
MicroRNA (miR)-148a-3p is most frequently upregulated in solid tumors, such as colorectal cancer (CRC). This study aimed to elucidate the role of miR-148a-3p in CRC cell proliferation and immune escape and its potential mechanism. miR-148a-3p and Kruppel-like transcription factor 4 (KLF4) expressions were quantified by western blot and quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), and immune evasion abilities of CRC cells were evaluated with the cell counting kit-8 assay, Transwell, western blot, and enzyme-linked immunosorbent assays. The proliferation or apoptosis of CD8+ and CD4+ T cells after coculture with CRC cells was assessed by flow cytometry. Dual-luciferase reporter gene testing was used to validate the targeting association between KLF4 and miR-148a-3p. A nude mouse subcutaneous graft tumor model was constructed, and CD8+ T cell infiltration was detected by immunohistochemistry and flow cytometry. miR-148a-3p exhibited a high level, while KLF4 was under-expressed in CRC cells; miR-148a-3p negatively regulated the KLF4 level. Overexpression of miR-148a-3p enhanced CRC cell proliferation, migration, invasion, EMT, and immune escape; silencing miR-148a-3p caused the opposite trend; moreover, the said biological functions of CRC cells were weakened with overexpression of KLF4 but enhanced with silencing of KLF4; silencing KLF4 weakened the influences of dampened miR-148a-3p on CRC development. Silencing miR-148a-3p promoted the infiltration of CD8+ T cells and inhibited tumor growth. In summary, miR-148a-3p promotes CRC cell proliferation and immune evasion by regulating the expression of KLF4. This finding can be used for reference when developing a new way of CRC treatment.
微小RNA（miR）-148a-3p在实体瘤（如结直肠癌，CRC）中最常上调。本研究旨在阐明miR-148a-3p在CRC细胞增殖和免疫逃逸中的作用及其潜在机制。通过蛋白质免疫印迹法和定量实时聚合酶链反应（qRT-PCR）对miR-148a-3p和 Kruppel样转录因子4（KLF4）的表达进行定量分析。采用细胞计数试剂盒-8法、Transwell小室实验、蛋白质免疫印迹法和酶联免疫吸附测定法评估CRC细胞的增殖、迁移、侵袭、上皮-间质转化（EMT）和免疫逃逸能力。通过流式细胞术评估CD8⁺和CD4⁺T细胞与CRC细胞共培养后的增殖或凋亡情况。采用双荧光素酶报告基因检测验证KLF4与miR-148a-3p之间的靶向关系。构建裸鼠皮下移植瘤模型，通过免疫组织化学和流式细胞术检测CD8⁺T细胞浸润情况。miR-148a-3p在CRC细胞中呈高水平表达，而KLF4表达下调；miR-148a-3p负向调节KLF4水平。miR-148a-3p过表达增强了CRC细胞的增殖、迁移、侵袭、EMT和免疫逃逸能力；沉默miR-148a-3p则产生相反的趋势；此外，CRC细胞的上述生物学功能随KLF4过表达而减弱，但随KLF4沉默而增强；沉默KLF4减弱了miR-148a-3p抑制对CRC发展的影响。沉默miR-148a-3p促进了CD8⁺T细胞浸润并抑制肿瘤生长。综上所述，miR-148a-3p通过调节KLF4的表达促进CRC细胞增殖和免疫逃逸。这一发现可为开发新的CRC治疗方法提供参考。
