江苏大学学报（医学版）2025，Vol.35Issue(2)：145-153,9.DOI:10.13312/j.issn.1671-7783.y240075
Circ_0022382/let-7a-5p/PI3K/AKT/mTOR轴促进乳腺癌细胞增殖和迁移
Circ_0022382/let-7a-5p/PI3K/AKT/mTOR axis promotes the proliferation and migration of breast cancer cells
刘伟 1张俊 2张佳雯 1叶雨 1诸健琴 1虞绮雯 1李涛 3孙晓春 1陈华标3
作者信息
- 1. 江苏大学医学院,江苏 镇江 212013
- 2. 扬州大学附属医院检验科,江苏 扬州 225012
- 3. 宁波大学医学院,浙江 宁波 315211
- 折叠
摘要
Abstract
Objective:To study the effects of circ_0022382 on the proliferation and migration of breast cancer cells and the possible underlying mechanism.Methods:①Screening for circRNAs that are highly expressed in breast cancer.Firstly,the differential analysis of the circRNA-related GEO dataset(GSE165884)and miRNA-related GEO dataset(GSE45498)of breast cancer were carried out,respectively,and then the miRNAs bound by the differentially expressed circRNAs were predicted by the ENCORI database,and finally the intersecting miRNAs and the miRNAs predicted by the GEO database were obtained by the Venn diagram.② The complementary DNA(cDNA)and genomic DNA(gDNA)derived from MDA-MB-231 of breast cancer cells were PCR with convergent primers and divergent primers,respectively,and the PCR products were separated by agarose gel electrophoresis to verify the circular structure of the circ_0022382.③ Real-time quantitative PCR(qRT-PCR)was used to detect circ_0022382 expression in breast cancer cells MDA-MB-231 and MCF-7 and normal breast epithelial cell MCF-10A,as well as in breast cancer tissues and adjacent tissues,respectively.④ MDA-MB-231 cells and MCF-7 cells were transfected with si-NC and si-circ_0022382,qRT-PCR was used to detect transfection efficiency,clone formation assay and scratch healing assay were used to detect the proliferation and migration of breast cancer cells.qRT-PCR was used to detect the expression level of let-7a-5p.The si-circ_0022382 group was transfected with let-7a-5p inhibitor NC and let-7a-5p inhibitor,respectively,and the proliferation and migration of breast cancer cells were detected by clone formation assay and scratch healing assay.⑤ The downstream signaling pathway of let-7a-5p was analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment.MDA-MB-231 cells and MCF-7 cells were transfected with let-7a-5p NC and let-7a-5p mimic,respectively,and the expression of PI3K/AKT/mTOR signaling pathway-related proteins was detected by Western blotting and cellular immunofluorescence,and the expression of p-AKT in breast cancer cells co-transfected with let-7a-5p inhibitor and si-circ_0022382 was detected by Western blotting.Results:① GSE165884 differential analysis yielded 37 high-expressing circRNAs,GSE45498 differential analysis yielded 6 low-expressing miRNAs,and among the 37 high-expression circRNAs,circ_0022382 corresponding miRNAs and 6 low-expressing miRNAs had intersecting gene let-7.② cDNA derived from breast cancer cell MDA-MB-231 can be amplified by both divergent primers and convergent primers,whereas gDNA can only be amplified by convergent primers.③ Compared with MCF-10A,circ_0022382 was significantly more expressed in MDA-MB-231 and MCF-7(P<0.001).The expression of circ_0022382 in breast cancer tissues was also higher than that in adjacent tissues(P<0.05).④ After knocking down circ_0022382,the proliferation and migration of MDA-MB-231 and MCF-7 cells decreased significantly(P<0.001),and the expression level of let-7a-5p increased significantly(P<0.01 and P<0.05).Co-transfection of let-7a-5p inhibitor significantly rescued the decreased proliferation and migration of MDA-MB-231 and MCF-7 cells transfected with si-circ_0022382(P<0.01 and P<0.001).⑤ The results of KEGG signaling pathway enrichment analysis showed that PI3K-AKT and mTOR signaling pathways were significantly enriched.Compared with the let-7a-5p NC group,the expression of p-AKT,p-PI3K and mTOR proteins in the let-7a-5p mimic group significantly decreased.Co-transfection of let-7a-5p inhibitor significantly rescued the decrease of p-AKT expression of MDA-MB-231 and MCF-7 cells transfected with si-circ_0022382.Conclusion:Circ_0022382 is highly expressed in breast cancer and promotes the proliferation and migration of breast cancer cells by targeting the let-7a-5p/PI3K/AKT/mTOR axis,which may be a potential target for breast cancer treatment and diagnosis.关键词
circ_0022382/let-7a-5p/PI3K/AKT/mTOR信号通路/增殖/迁移Key words
circ_0022382/let-7a-5p/PI3K/AKT/mTOR signaling pathways/proliferation/migration分类
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刘伟,张俊,张佳雯,叶雨,诸健琴,虞绮雯,李涛,孙晓春,陈华标..Circ_0022382/let-7a-5p/PI3K/AKT/mTOR轴促进乳腺癌细胞增殖和迁移[J].江苏大学学报（医学版）,2025,35(2):145-153,9.
