circRNA_001569通过miR-145/HBXIP轴影响乳腺癌细胞的恶性生物学行为

circRNA_001569 affects the malignant biological behavior of breast cancer cells through the miR-145/HBXIP axis

目的:探讨circRNA001569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circRNA001569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-circRNA001569、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,circRNA001569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circRNA001569和HBXIP之间的靶向关系;circRNA001569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:circRNA001569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。

Objective: To investigate the role of circRNA001569 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 circRNA001569 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-circRNA001569,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 circRNA001569 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,circRNA001569 and HBXIP. Over-expression of circRNA001569 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:circRNA001569 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.