[关键词]
[摘要]
目的:探讨 circRNA_001569 通过 miR-145/HBXIP 轴在乳腺癌细胞增殖、侵袭、迁移中发挥的作用。方法:收集2016年1月至2019年1月期间衡水市人民医院收治的30例乳腺癌患者的癌组织和癌旁组织。qPCR检测circRNA_001569在乳腺癌组织、癌旁组织以及细胞系中的表达。生物信息学工具预测miR-145的靶基因,RNA免疫沉淀(RNA immunoprecipitation,RIP)和双荧光素酶报告基因实验检测 miR-145 或靶基因之间的相互作用 ;向乳 腺 癌 MDA-MB-231 和 MCF-7细胞中转染si-circRNA_001569、miR-145 mimics或miR-145 inhibitor,建立基因过表达或沉默的细胞模型,qPCR和Western blotting分别检测转染对相关基因和蛋白表达的影响,CCK-8法、Transwell实验检测转染对细胞增殖、侵袭和迁移的影响。结果:在乳腺癌组织和乳腺癌细胞中,circRNA_001569 和 HBXIP 均呈高表达、miR-145 呈低表达。RIP 分析和双荧光素酶实验证实了 miR-145 与circRNA_001569和HBXIP之间的靶向关系;circRNA_001569或HBXIP过表达促进MDA-MB-231和MCF-7细胞的增殖、侵袭和迁移(均 P<0.01),而 miR-145 过表达起相反的作用(均 P<0.01)。结论:circRNA_001569 可能通过下调 miR-145 的表达、上调HBXIP的表达从而促进乳腺癌细胞的增殖、侵袭和迁移。
[Key word]
[Abstract]
Objective: To investigate the role of circRNA_001569 in the proliferation, invasion and migration of breast cancer cells via miR-145/HBXIP axis. Methods: Collected 30 cases of cancerous tissue and 30 adjacent tissues of breast cancer patients treated in the Hengshui People's Hospital from January 2016 to January 2019. qPCR detects the expression of circRNA_001569 in breast cancer tissues, adjacent tissues and cell lines. Bioinformatics tools were used to predict the target genes of miR-145. RNA immunoprecipitation (RIP) and Dual luciferase reporter gene assay were used to detect the interaction between genes or proteins. si-circRNA_001569,miR-145 mimics or miR-145 inhibitor were transfected into breast cancer MDA-MB-231 and MCF-7 cells to establish gene overexpression or silence cell models. qPCR and Western blotting were used to detect the expressions of related genes and proteins.CCK-8 method and Transwell test were used to detect the proliferation, invasion and migration of transfected cells. Results: In breast cancer tissues and breast cancer cells, the expressions of circRNA_001569 and HBXIP were up-regulated, while the expression of miR-145 was down-regulated; RIP analysis and Dual luciferase analysis revealed the targeting relationship among miR-145,circRNA_001569 and HBXIP. Over-expression of circRNA_001569 or HBXIP promoted the proliferation, invasion and migration of MDA-MB-231 and MCF-7 cells (all P<0.01), while miR-145 overexpression had the opposite effect (all P<0.01). Conclusion:circRNA_001569 may promote the proliferation, invasion and migration of breast cancer cells by down-regulating the expression of miR-145 and up-regulating the expression of HBXIP.
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[基金项目]
河北省重点研发计划资助项目(No.19277799D)