hsa_circ_0070260靶向miR-27a-3p/Fbxw7对乳腺癌增殖、侵袭调控研究

Hsa_circ_0070260/miR-27a-3p/Fbxw7 regulates the proliferation and migration of breast cancer cells

目的探讨hsa_circ_0070260对乳腺癌增殖、侵袭的调控作用并揭示相关机制。方法利用RT-qPCR技术检测hsa_circ_0070260在乳腺癌细胞系和对应的乳腺上皮细胞中表达情况。构建hsa_circ_0070260和Fbxw7荧光素酶报告实验载体,利用双荧光素酶报告系统检测hsa_circ_0070260、miR-27a-3p和Fbxw7之间的靶向调控关系。构建hsa_circ_0070260干扰载体,miR-27a-3p干扰/过表达载体,Fbxw7过表达载体,转染MDA-MB-231细胞后利用平板克隆实验、Transwell实验技术检测hsa_circ_0070260、miR-27a-3p和Fbxw7之间的靶向作用对细胞增殖和迁移的调控作用。结果结果显示相对正常的MCF-10A细胞,hsa_circ_0070260在乳腺癌细胞系MCF-7、T47D、MDA-MB-231、BT-20细胞中表达上调,其中MDA-MB-231细胞中表达上调最为显著。利用双荧光素酶报告系统检测发现hsa_circ_0070260和miR-27a-3p、miR-27a-3p和Fbxw7之间存在靶向作用关系。利用MDA-MB-231细胞研究发现下调hsa_circ_0070260后抑制了细胞增殖和迁移;在hsa_circ_0070260敲低基础上干扰miR-27a-3p后恢复了细胞增殖、迁移能力;在hsa_circ_0070260敲低基础上过表达Fbxw7后同样恢复了细胞增殖、迁移能力,上述结果显示hsa_circ_0070260可能通过吸附miR-27a-3p来促进Fbxw7表达。进一步实验研究发现miR-27a-3p过表达抑制了细胞增殖、迁移,而miR-27a-3p过表达后上调Fbxw7恢复了细胞增殖、迁移能力,说明miR-27a-3p具有抗肿瘤作用,且Fbxw7是miR-27a-3p的下游靶点。结论hsa_circ_0070260通过吸附miR-27a-3p来促进Fbxw7表达是导致乳腺癌细胞增殖、迁移的重要原因,靶向hsa_circ_0070260对乳腺癌细胞增殖、迁移具有一定的抑制作用,希望通过该研究为乳腺癌的临床治疗提供一定的理论基础和治疗靶点。

Objective To explore the regulatory effect of hsa_circ_0070260 on proliferation and invasion of breast cancer cells and the relevant mechanism.Methods To investigate the regulation of hsa_circ_0070260 on proliferation and invasion of breast cancer cells and reveal the relevant mechanism.RT-qPCR was used to detect the expression of hsa_circ_0070260 in breast cancer cell lines and breast epithelial cells.Hsa_circ_0070260 and Fbxw7 luciferase reporting experimental vectors were constructed,and the targeted regulatory relationship between hsa_circ_0070260,miR-27a-3p and Fbxw7 was detected by dual-luciferin reporting experiment.Hsa_circ_0070260 interference vector,miR-27a-3p interference/over-expression vector,and Fbxw7 over-expression vector were constructed.After transfection in MDA-MB-231 cells,the targeted effects of hsa_circ_0070260,miR-27a-3p and Fbxw7 on cell proliferation and migration were detected by the transwell assay.Results The results showed that compared with normal MCF-10A cells,hsa_circ_0070260 was up-regulated in breast cancer cell lines MCF-7,T47D,MDA-MB-231,and BT-20 cells,among which MDA-MB-231 cells were the most significantly upregulated.The dual-luciferin report assay revealed a targeting relationship between hsa_circ_0070260 and miR-27a-3p,miR-27a-3p and Fbxw7.In MDA-MB-231 cells,it was found that down-regulation of hsa_circ_0070260 inhibited cell proliferation and migration.The proliferation and migration of miR-27a-3p cells were restored after hsa_circ_0070260 knockdown.Overexpression of Fbxw7 on the basis of hsa_circ_0070260 knockdown also restored the proliferation and migration ability of cells.It was speculated that hsa_circ_0070260 might promote the expression of Fbxw7 by adsorbing miR-27a-3p.Further experimental found that the overexpression of miR-27a-3p inhibited the proliferation and migration of cells,while the upregulation of Fbxw7 restored the proliferation and migration ability after the overexpression of miR-27a-3p,indicating that miR-27a-3p has anti-tumor effect,and Fbxw7 is the downstream target of miR-27a-3p.Conclusion Hsa_circ_0070260 promotes the expression of Fbxw7 by adsorbing miR-27a-3p,which is an important cue for the proliferation and migration of breast cancer cells.Targeted hsa_circ_0070260 has a certain inhibitory effect on the proliferation and migration of breast cancer cells.It is hoped that this study can provide a certain theoretical basis and therapeutic target for clinical treatment of breast cancer.