南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (8): 1790-1798.doi: 10.12122/j.issn.1673-4254.2026.08.07

• • 上一篇    

miR-183-5p通过靶向调控KLHL24影响三阴性乳腺癌细胞增殖、侵袭、迁移及铁死亡

王坤1(), 侯顺福2, 李勇1, 刘清华3, 尹崇高4, 李洪利1,2()   

  1. 1.山东第二医科大学，医学研究实验中心，山东 潍坊 261053
    2.山东第二医科大学，基础医学院病理学教研室，山东 潍坊 261053
    3.山东第二医科大学，人体解剖学教研室，山东 潍坊 261053
    4.山东第二医科大学，护理学院，山东 潍坊 261053
  • 收稿日期:2026-01-16 出版日期:2026-08-20 发布日期:2026-08-01
  • 通讯作者: 李洪利 E-mail:wangkun8341@sdsmu.edu.cn;lihongli@sdsmu.edu.cn
  • 作者简介:王 坤，助理实验师，硕士，E-mail: wangkun8341@sdsmu.edu.cn
  • 基金资助:
    国家自然科学基金(82373043);山东省自然科学基金(ZR2022MH311);潍坊市科学技术发展计划项目(2023YX037)

Overexpression of miR-183-5p enhances malignant phenotype and inhibits ferroptosis of triple-negative breast cancer cells by negatively regulating KLHL24

Kun WANG1(), Shunfu HOU2, Yong LI1, Qinghua LIU3, Chonggao YIN4, Hongli LI1,2()   

  1. 1.Medical Research Center, Shandong Second Medical University, Weifang 261053, China
    2.Department of Clinical Pathology, Shandong Second Medical University, Weifang 261053, China
    3.Human Anatomy Teaching and Research Section, Shandong Second Medical University, Weifang 261053, China
    4.College of Nursing, Shandong Second Medical University, Weifang 261053, China
  • Received:2026-01-16 Online:2026-08-20 Published:2026-08-01
  • Contact: Hongli LI E-mail:wangkun8341@sdsmu.edu.cn;lihongli@sdsmu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(82373043)

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摘要：

目的 探究miR-183-5p靶向调控Kelch样蛋白24（KLHL24）对三阴性乳腺癌（TNBC）细胞增殖、侵袭、迁移及铁死亡影响的研究。 方法 应用生物信息学方法对下载的有关乳腺癌的数据集和转录组数据进行差异化分析，筛选出目标基因；再结合多个在线数据库预测并确定目标基因的靶基因。采用RT-qPCR验证目标基因在正常乳腺上皮细胞和乳腺癌细胞系中的表达情况。通过双荧光素酶报告基因实验验证目标基因与靶基因的靶向关系。通过Western blotting检测转染目标基因过表达质粒后细胞中靶基因编码蛋白表达变化。将MDA-MB-231细胞分为4组并转染：NC-miR + NC-KLHL24组（miR-183-5p对照质粒+KLHL24对照质粒共转染）、oe-miR-183-5p+ NC-KLHL24组（miR-183-5p过表达质粒+KLHL24对照质粒共转染）、NC-miR +oe-KLHL24组（miR-183-5p对照质粒+KLHL24过表达质粒共转染）、oe-miR-183-5p+oe-KLHL24组（miR-183-5p过表达质粒+KLHL24过表达质粒共转染）。使用EdU、Transwell及划痕愈合实验评估细胞增殖、侵袭和迁移能力变化，采用Western blotting技术，分析铁死亡核心调控通路（GPX4、SLC7A11、Nrf2）相关蛋白的相对表达水平，活性氧（ROS）实验、脂质过氧化物（LPO）实验、丙二醛（MDA）实验和还原型谷胱甘肽（GSH））实验检测目标基因以及其靶基因对上述铁死亡指标的影响。 结果 miR-183-5p在乳腺癌组织中的表达量高于正常乳腺组织（P<0.001），在乳腺癌细胞系中表达亦均高于正常乳腺上皮细胞，其中在MDA-MB-231细胞中表达最高（P<0.0001）。双荧光素酶报告基因实验证实miR-183-5p可以与KLHL24的3'-UTR靶向结合（P<0.001）。Western blotting显示过表达miR-183-5p后可降低靶蛋白KLHL24的表达（P<0.0001）。EdU细胞增殖、Transwell侵袭和划痕愈合实验结果显示，在TNBC细胞中过表达miR-183-5p后能明显促进细胞的增殖、侵袭和迁移能力（P<0.001），过表达KLHL24可以抵消miR-183-5p的促进作用。过表达miR-183-5p可上调GPX4、SLC7A11、Nrf2蛋白表达（P<0.05），过表达KLHL24则下调上述蛋白表达（P<0.05），且共表达KLHL24可逆转miR-183-5p对这些蛋白的上调效应。铁死亡指标检测结果表明，过表达miR-183-5p后可以下调细胞内ROS、LPO和MDA含量（P<0.0001），上调GSH的含量（P<0.0001），过表达KLHL24则能有效逆转miR-183-5p对上述铁死亡相关指标的调控作用。 结论 miR-183-5p可以通过负向调控KLHL24增强TNBC细胞增殖、侵袭和迁移能力，并抑制铁死亡的发生。

关键词: 三阴性乳腺癌, miR-183-5p, KLHL24, 增殖, 侵袭和迁移, 铁死亡

Abstract:

Objective To investigate the effects of miR-183-5p overexpression on proliferation, invasion, migration, and ferroptosis of triple-negative breast cancer (TNBC) cells and the role of Kelch-like protein 24 (KLHL24) in mediating these effects. Methods Bioinformatics analyses were conducted to screen differentially expressed genes in breast cancer (BRCA) and predict the target gene of miR-183-5p. RT-qPCR was used to verify miR-183-5p expression in normal breast epithelial and TNBC cell lines. Dual-luciferase reporter assay was performed to confirm direct binding of miR-183-5p to 3'-UTR of KLHL24. Western blotting was used to detect KLHL24 expression in different TNBC cells after transfection with miR-183-5p-overexpressing plasmids. MDA-MB-231 cells were co-transfected with NC-miR and NC-KLHL24, oe-miR-183-5p and NC-KLHL24, NC-miR and oe-KLHL24, or oe-miR-183-5p and oe-KLHL24 plasmids, and the changes in cell proliferation, migration and invasion were examined using EdU, Transwell, and wound healing assays. Western blotting was used to detect the changes in expressions of ferroptosis-related proteins (GPX4, SLC7A11, and Nrf2), and ROS, LPO, MDA, and GSH levels in the transfected cells were determined. Results The expression of miR-183-5p was significantly elevated in BRCA tissues and TNBC cell lines, and MDA-MB-231 cells showed the highest miR-183-5p expression. Dual-luciferase reporter assay confirmed the direct binding of miR-183-5p to 3'-UTR of KLHL24. Overexpression of miR-183-5p significantly downregulated KLHL24 expression. In TNBC cells, miR-183-5p overexpression obviously promoted cell proliferation, invasion, and migration, upregulated the expressions of GPX4, SLC7A11 and Nrf2, reduced the levels of ROS, LPO and MDA, and increased the level of GSH (P<0.0001); these effects were significantly reversed by co-transfection of the cells with KLHL24-overexpressing plasmids. Conclusion Overexpression of miR-183-5p enhances TNBC cell proliferation, invasion, migration and inhibits ferroptosis by negatively regulating KLHL24.

Key words: triple-negative breast cancer, miR-183-5p, KLHL24, proliferation, invasion and migration, ferroptosis