miR-597调控FOSL2抑制乳腺癌细胞增殖、迁移及侵袭的研究

miR-597 inhibits breast cancer cell proliferation,migration and invasion through FOSL2

目的探讨miR-597对乳腺癌细胞增殖、迁移及侵袭的影响,及其与FOS样抗原2(Fos-related antigen 2,FOSL2)的靶向关系。方法选取乳腺癌细胞株T-47D,SK-BR-3,MDA-MB-231,MCF7及BT-474,正常乳腺细胞株MCF10A为研究对象,分别转染miR-597(miR-597组)及RNA阴性对照序列(NC组),同时选取30例乳腺癌病人癌组织及癌旁正常乳腺组织,迅速液氮冻存以提取RNA用于qRT-PCR检测。采用实时荧光定量PCR法检测细胞、组织中miR-597表达水平,MTT、流式细胞术检测细胞增殖能力,Transwell小室实验检测细胞迁移、侵袭能力,Western blot检测细胞FOSL2表达水平,生物信息预测miR-597与FOSL2的靶向作用关系,荧光素酶实验验证。结果乳腺癌组织miR-597水平低于正常乳腺组织,乳腺癌细胞株细胞miR-597表达低于正常乳腺细胞株细胞,差异有统计学意义(P<0.01),miR-597组细胞增殖能力明显低于NC组,差异有统计学意义(t=9.833,P=0.003),miR-597组G0G1期细胞百分率高于NC组,S期细胞百分率低于NC组(t=10.215,10.336,P=0.000,0.000),miR-597组细胞侵袭能力低于NC组(t=8.404,P=0.000),miR-597组细胞迁移能力低于NC组(t=5.801,P=0.016),miR-597组细胞FOSL2表达水平低于NC组(t=13.658,P=0.000),miR-597与FOSL2有良好的靶向关系。结论miR-597在乳腺癌组织及细胞株中低表达,其高表达可可抑制乳腺癌细胞的增殖、迁移及侵袭能力,其机制与调控FOSL2有关。

Objective To investigate the effect of miR-597 on the proliferation,migration and invasion of breast cancer cells,and its relationship with FOS related antigen 2( FOSL2). Methods Breast cancer cell lines T-47 D,SK-BR-3,MDA-MB-231,MCF7,BT-474 and normal breast cell line MCF10 A were selected as the study objects. According to the transfection,it was divided into: NC group( null plasmid) and miR-597 group( simulated sequence). Meanwhile,30 patients with breast cancer tissue and normal breast tissue adjacent to cancer were selected,rapid cryopreservation of liquid nitrogen to extract RNA for qRT-PCR. The expression level of miR-597 in cells and tissues was detected by real-time fluorescence quantitative PCR. The cell proliferation was detected by MTT and flow cytometry. The cell migration and invasion were detected by Transwell chamber. The expression of FOSL2 was detected by Western blot. The target effect of miR-597 and FOSL2 was confirmed by bioinformatics methodsand the luciferase reporter assay. Results The expression of miR-597 in breast cancer tissue was lower than that in normal breast tissue( P < 0. 01). The cell proliferation ability of miR-597 in breast cancer cell line was lower than that in normal breast cell line( P < 0. 01). The percentage of G0 G1 phase cells in miR-597 group was higher than that in NC group,and the percentage of S-phase cells was lower than that in NC group( t = 10. 215,10. 336,P = 0. 000,0. 000). The cell invasion ability of miR-597 group was lower than that of NC group( t = 8. 404,P = 0. 000). The cell migration ability of miR-597 group was lower than that of NC group( t = 5. 801,P = 0. 016). The expression level of FOSL2 in miR-597 group was lower than that of NC group( t = 13. 658,P = 0. 000). MiR-597 had a good targeting relationship with FOSL2. Conclusion miR-597 was low expressed in breast cancer tissue and cell lines. The high expression of miR-597 can inhibit the proliferation,migration and invasion of breast cancer cells,and which was related to the regulation of FOSL2.