%0 Journal Article %T MiR-182/FOXF2信号通路在三阴性乳腺癌侵袭及血管生成中的意义 %A 于金玲 %A 沈卫达 %A 高蓓敏 %A 赵海燕 %A 曹小曼 %J 同济大学学报(医学版) %D 2018 %R 10.16118/j.1008-0392.2018.01.011 %X 目的 探讨miR-182对三阴性乳腺癌侵袭的影响及其与靶基因FOXF2的调控关系。方法 通过CCK-8增殖实验和Transwell实验进行体外细胞功能验证，检测miR-182对乳腺癌细胞增殖和侵袭的影响；采用Targetscan软件预测miR-182靶向调控FOXF2基因表达结果；通过靶点验证及靶点突变测试验证miR-182在基因FOXF2上结合位点；采用qRT-PCR检测miR-182对FOXF2基因表达的靶向调控，ELISA检测HUVEC细胞系中VEGF蛋白表达。结果体外细胞功能实验证明，miR-182可明显促进乳腺癌细胞的增殖与侵袭（P＜0.01）；荧光素酶报告基因实验证实，miR-182可通过结合FOXF2基因3′-非翻译区688～695位点（3′-untranslational region, 3′-UTR）而抑制其表达；qRT-PCR检测发现，转染miR-182后MCF-7中FOXF2表达量明显降低(P＜0.01)；体外血管生成实验表明，转染miR-182后HUVEC细胞培养液上清液中VEGF蛋白含量显著升高（P=0.004)。结论 miR-182作为促癌因素，直接作用于FOXF2基因，引起该基因表达下调，促进血管生成，发挥侵袭作用，可能是促进三阴性乳腺癌转移的生物学因素机制之一。</br>Objective To investigate the effects of miR-182 on proliferation and invasion ability of three-negative breast cancer (TNBC) and its relation to target gene FOXF2. Methods The migration and proliferation of breast cancer MDA-MB-231 (high invasion) and MCF7 cells (low invasion) were evaluated with Transwell assay and CCK-8 assay, respectively. The candidate microRNAs targeting FOXF2 were searched with bioinformatic prediction software Targetscan and verified by luciferase assay using FOXF2 3′-untranslated region（3′-UTR）reporter. The MDA-MB-231 cells and MCF7 cells were transfected with antagomiR-182 and miR-182 mimics, respectively. The expression of FOXF2 was detected with real-time quantitative PCR（qRT-PCR）in MDA-MB-231 and MCF7 cells after transfection. The expression of VEGF protein in HUVEC cells was detected by ELISA method. Results The migration and invasive ability of MCF-7 cells was significantly increased after transfection of miR-182 mimics (P＜0.01). FOXF2 at position 688-695 was found as the potential target region of miR-182. The luciferase reporter assay verified the predicted results. The qRT-PCR revealed the over-expression of miR-182 with the reduced FOXF2 mRNA expression in MCF-7 cells after transfection of miR-182 mimics（P＜0.01）．The contents of VEGF protein in HUVEC cell culture media were significantly increased after transfection of miR-182 (P= 0.004). Conclusion miR-182 promotes angiogenesis, proliferation and migration of triple negative breast cancer, which is associated with down-regulation of its target gene FOXF2 expression %K 三阴性乳腺癌 miR-182 FOXF2 侵袭 血管生成< %K /br> %K triple-negative breast cancer microRNA-182 FOXF2 migration angiogenesis %U http://tjyxxb.cnjournals.cn/ch/reader/view_abstract.aspx?file_no=20180111&flag=1