南方医科大学学报2025，Vol.45Issue(6)：1200-1211,12.DOI:10.12122/j.issn.1673-4254.2025.06.09
敲除CDC20可明显抑制宫颈癌细胞的增殖及侵袭转移
CRISPR-Cas9-mediated CDC20 gene knockout inhibits cervical cancer cell proliferation,invasion and metastasis
摘要
Abstract
Objective To study the effect of CDC20 knockdown on proliferation,migration and invasion of cervical cancer cells and its underlying mechanism.Methods CDC20 expression in cervical cancer tissues was analyzed using the TCGA database,and the protein expressions of CDC20 and β-Catenin in clinical specimens of cervical cancer and adjacent tissues were detected using immunohistochemistry.A dual target sgRNA2&7 sequence for CDC20 gene was designed for CDC20 gene knockdown in cervical cancer C33A cells using CRISPR/Cas9 technology,and CDC20 mRNA and protein expression levels in the transfected cells were detected using qRT-PCR and Western blotting.The changes in proliferation,cell cycle,apoptosis,migration and invasiveness of the transfected cells were evaluated using colony-forming assay,fluorescence activated cell sorting(FACS)and Transwell assay.In the animal experiment,naïve C33A cells and the cells with CDC20 knockdown were injected subcutaneously into the left and right axillae of nude mice(n=5)to observe tumor growth.The expressions of CDC20 and β-Catenin proteins in transfected cells and the xenograft were analyzed using Western blotting,and their interaction was confirmed by co-immunoprecipitation(CoIP)and immunofluorescence co-localization assays.Results Cervical cancer tissues expressed significantly higher CDC20 and β-Catenin levels than the adjacent tissues.C33A cells with CDC20 knockdown showed reduced proliferation,increased apoptosis,and lowered migration and invasion abilities.CDC20 knockdown significantly suppressed the growth of C33A cell xenograft in nude mice,and the tumor-bearing mice did not exhibit obvious body mass changes.CDC20 and β-Catenin levels were both significantly lowered in C33A cells with CDC20 knockdown.Co-immunoprecipitation and co-localization assays confirmed the interaction between CDC20 and β-Catenin.Conclusion CDC20 is highly expressed in cervical cancer tissues,and CDC20 knockdown can suppress proliferation,invasion,and metastasis while enhancing apoptosis of C33A cells,which is closely related with the regulation of the Wnt/β-Catenin signaling pathway.关键词
CRISPR/Cas9/CDC20/宫颈癌/Wnt/β-CateninKey words
CRISPR/Cas 9/CDC20/cervical cancer/Wnt/β-Catenin引用本文复制引用
莫艳秀,舒洋,莫钰兰,刘峻彤,徐欧欧,邓华菲,王岐本..敲除CDC20可明显抑制宫颈癌细胞的增殖及侵袭转移[J].南方医科大学学报,2025,45(6):1200-1211,12.基金项目
湖南省教育厅资助科研项目(23A0589) （23A0589）
湘南学院高层次人才科研启动基金项目(2021-A20) （2021-A20）
湖南省大学生创新训练计划项目(湘教通[2024]191号,4854) （湘教通[2024]191号,4854）
