赣南医科大学学报2025，Vol.45Issue(12)：1138-1145,8.DOI:10.3969/j.issn.2097-7174.2025.12.003
circ-CCDC66促进MCF-7细胞增殖、迁移、侵袭及干细胞样表型形成
circ-CCDC66 in promoting proliferation,migration,invasion and stem-like phenotype in MCF-7 cells
摘要
Abstract
Objective:To clarify the expression pattern and biological role of circular RNA circ-CCDC66 in breast cancer,and to preliminarily explore its potential downstream target genes.Methods:Normal breast epithelial cells(MCF-10A)and four breast cancer cell lines(MCF-7,MDA-MB-231,MDA-MB-453,and MDA-MB-468)were selected.The expression of circ-CCDC66 was examined by quantitative PCR(qPCR),and its circular structure was validated using agarose gel electrophoresis and Sanger sequencing.The circ-CCDC66 overexpression plasmid or circ-CCDC66 siRNAs were transfected into MCF-7 cells by Lipofectamine 2000 to construct the control group and the experimental group.CCK-8 assays were performed to assess cell viability,Transwell assays were used to evaluate migratory and invasive abilities,and mammosphere formation assays were conducted to examine stem cell-like phenotypes.The expression of circ-CCDC66 was confirmed using qPCR in clinical tissue samples(including cancer tissues and adjacent tissues).RNA sequencing was used to identify differentially expressed mRNAs,and enrichment pathway analysis was employed to identify key metabolic signaling events.Two representative genes,BPNT1 and KYAT1,were selected for further qPCR validation.Results:circ-CCDC66 was found to be upregulated in breast cancer cells and exhibited a stable circular configuration.Its overexpression enhanced the proliferation,migration,invasion,and stem-like properties of MCF-7 cells(P＜0.05).Besides,siRNA interference experiments further verified the functional role of circ-CCDC66 in breast cancer cells.Transcriptome analysis of cells in the PCDH-NC and PCDH-circ-CCDC66 groups revealed 61 differentially expressed genes,including 34 upregulated and 27 downregulated,which were predominantly enriched in sulfur metabolism,selenocompound metabolism,and cyanoamino acid metabolism pathways.qPCR validation demonstrated that BPNT1 and KYAT1 were downregulated in the circ-CCDC66 overexpression group,consistent with the RNA-seq results.Conclusion:circ-CCDC66 enhances proliferation,migration,invasion,and mammosphere formation in MCF-7 cells,and downregulates the expression of BPNT1 and KYAT1.关键词
乳腺癌/circ-CCDC66/增殖/迁移/干细胞Key words
Breast cancer/circ-CCDC66/Proliferation/Migration/Stem cell分类
医药卫生引用本文复制引用
曾伟华,李微绽,黎颂铭,郭锦全,郭志云,唐杰东,邹国荣,苏珍,殷琛,何艳..circ-CCDC66促进MCF-7细胞增殖、迁移、侵袭及干细胞样表型形成[J].赣南医科大学学报,2025,45(12):1138-1145,8.基金项目
广州市科技计划项目(202201010779) （202201010779）
