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人源环状RNA CSPP1/微小RNA-361-5p/整合素β1通过调控PI3K-Akt信号通路调节宫颈癌的增殖和迁移

Hsa_circ_CSPP1/MiR-361-5p/ITGB1 Regulates Proliferation and Migration of Cervical Cancer (CC) by Modulating the PI3K-Akt Signaling Pathway.

作者信息

Yang Wenjie, Xie Tong

机构信息

Department of Gynecology, Jiangxi Maternal and Child Health Hospital, 318 Bayi Road, Donghu, Nanchang, 330006, Jiangxi, People's Republic of China.

Center of Health Management Physical Examination, Jiangxi Maternal and Child Health Hospital, 318 Bayi Road, Donghu, Nanchang, 330006, Jiangxi, People's Republic of China.

出版信息

Reprod Sci. 2020 Jan;27(1):132-144. doi: 10.1007/s43032-019-00008-5. Epub 2020 Jan 1.

Abstract

This study aimed to investigate the regulatory mechanism of circular RNA CSPP1 (hsa_circ_CSPP1) in cervical cancer. Based on GEO database, differentially expressed circRNAs and mRNAs related to cervical cancer were screened out by R software. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) term analysis were performed to analyze the functional and pathway enrichment of identified DEGs. In addition, Cytoscape software was used to build interaction network of DEGs. The mRNA expressions were examined by qRT-PCR. Western blot was conducted to view the expression of proteins. Cell proliferation and apoptosis were respectively evaluated using CCK-8 assay and flow cytometry, whereas cell migration abilities were detected by Transwell assay. The relationship among factors was validated by dual-luciferase reporter gene assay. The influence in cervical tumor growth was further verified through nude mouse model in vivo. Hsa_circ_CSPP1 and ITGB1 were high-expressed in cervical cancer, while miR-361-5p was low-expressed. Hsa_circ_CSPP1 knockdown or miR-361-5p overexpression could suppress cervical cancer cell proliferation and migration, whereas promoted cell apoptosis. In addition, further experiments demonstrated that both hsa_circ_CSPP1 and ITGB1 mRNA were targets of miR-361-5p. Repressing hsa_circ_CSPP1 restrained cell viability and mobility and induced apoptosis through sponging miR-361-5p. Meanwhile, miR-361-5p also inhibited cervical cancer tumorigenesis via downregulation of ITGB1. Knockdown of hsa_circ_CSPP1 impeded tumor growth through suppressing the expression of downstream gene ITGB1, PI3K, and Akt. Circular RNA hsa_circ_CSPP1 regulates cell migration and proliferation in cervical cancer through miR-361-5p/ITGB1 in PI3K-Akt signaling pathway.

摘要

本研究旨在探讨环状RNA CSPP1(hsa_circ_CSPP1)在宫颈癌中的调控机制。基于GEO数据库,利用R软件筛选出与宫颈癌相关的差异表达环状RNA和mRNA。进行京都基因与基因组百科全书(KEGG)通路分析和基因本体(GO)术语分析,以分析已鉴定的差异表达基因(DEG)的功能和通路富集情况。此外,使用Cytoscape软件构建DEG的相互作用网络。通过qRT-PCR检测mRNA表达。进行蛋白质免疫印迹法以观察蛋白质表达。分别使用CCK-8法和流式细胞术评估细胞增殖和凋亡,而通过Transwell实验检测细胞迁移能力。通过双荧光素酶报告基因实验验证各因素之间的关系。通过体内裸鼠模型进一步验证对宫颈肿瘤生长的影响。Hsa_circ_CSPP1和ITGB1在宫颈癌中高表达,而miR-361-5p低表达。敲低Hsa_circ_CSPP1或过表达miR-361-5p可抑制宫颈癌细胞增殖和迁移,同时促进细胞凋亡。此外,进一步实验表明Hsa_circ_CSPP1和ITGB1 mRNA均为miR-361-5p的靶标。抑制Hsa_circ_CSPP1通过海绵化miR-361-5p来抑制细胞活力和迁移并诱导凋亡。同时,miR-361-5p还通过下调ITGB1抑制宫颈癌肿瘤发生。敲低Hsa_circ_CSPP1通过抑制下游基因ITGB1、PI3K和Akt的表达来阻碍肿瘤生长。环状RNA hsa_circ_CSPP1通过PI3K-Akt信号通路中的miR-361-5p/ITGB1调节宫颈癌中的细胞迁移和增殖。

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