miR-411在乳腺癌中的表达及对肿瘤增殖转移的影响和机制研究  

The expression of miR-411 and its effects and mechanism research on proliferation and metastasis in breast cancer

作　　者：邓晓芳[1] 柳子川[1] 胡利娟[1] 胡小山[1] 徐岗[2] 

机构地区：[1]广州医科大学附属肿瘤医院内二科,广州510095 [2]广州医科大学附属肿瘤医院胸外科,广州510095

出　　处：《中国医师杂志》2017年第2期201-206,共6页Journal of Chinese Physician

基　　金：广东省医学科学技术研究基金项目（B2016070）;广州医科大学研究项目（2015C42）

摘　　要：目的探讨miR-411在乳腺癌中的表达及对肿瘤增殖转移的影响及部分分子机制。方法采用RT—PCR检测乳腺癌细胞和组织中miR-411表达情况。MTT、克隆形成实验和Transwell实验分别检测过表达和低表达miR-411对乳腺癌细胞增殖、迁移和侵袭的影响。RT—PCR和Western blot检测miR-41l对乳腺癌细胞生长因子受体结合蛋白2（GRB2）表达的影响，双荧光素酶报告实验检测miR-411对GRB2的靶向作用。MTT、克隆形成实验和Transwell实验检测抑制GRB2的表达对乳腺癌细胞增殖和侵袭的影响。Westernblot检测高表达miR-411对乳腺癌细胞GRB2下游相关信号通路分子表达的影响。结果与癌旁组织相比，乳腺癌组织中miR-411表达明显减少（P〈0．05）。乳腺癌细胞SK—BR-3、BT-549、MDA—MB-231中miR-411呈低表达状态（P〈0．05）。与阴性对照组相比，转染miR-411 mimic可抑制乳腺癌细胞MDA—MB-231增殖、迁移和侵袭（P〈0．05）。Targetscan显示miR-411可靶向结合在GRB23’UTR 741—747位点，与阴性对照组相比，GRB2 3’UTR野生型质粒与miR-411共同转染可降低荧光活性（P〈0．05）。转染siGRB2可显著减少乳腺癌细胞MDA—MB-231中GRB2蛋白表达（P〈0．05），与阴性对照组相比，抑制GRB2的表达可减少乳腺癌细胞MDA—MB-231的增殖和克隆形成数（P〈0．05），Transwell实验表明，转染siGRB2可显著减少乳腺癌细胞MDA-MB-231的侵袭细胞数（P〈0．05）。结论miR-411与乳腺癌的发生发展密切相关，miR-411-GRB2-Ras轴有望成为乳腺癌生物治疗的新靶点。[ Abstract] Objective To investigate the relationship between miR-411 and breast cancer, and the expression of miR411 and its effects and mechanism research on proliferation and metastasis in breast cancer. Methods Real-time polymerase chain reaction （RT-PCR） was used to detect the expression of miR- 411 in breast cancer cells and tissues. Methyl thiazolyl tetrazolium （ MTT） , clone formation assay and Transwell assay were used to detect the effect of miR411 expression on the proliferation, migration and invasion in breast cancer cells. The effect of miR411 on growth factor receptor-bound protein 2 （GRB2） expression in breast cancer cells was detected by RT-PCR and Western blot. The direct effect of miR411 target on GRB2 was detected by dual luciferase reporter assay. MTT, clone formation assay and Transwell assay de- tect the effect of GRB2 expression on the proliferation and invasion in breast cancer ceils. Detection the effect high expression of miR411 on GRB2 downstream signaling pathway related molecules expression in breast cancer cell with Western blot. Results The expression of miR-411 in breast cancer tissues was significantly lower than that in adjacent non-cancerous tissues （P 〈 0.05 ）. The expression of miR-411 in breast cancer cells SK-BR-3, BT-549 and MDA-MB-231 was significantly lower （P 〈 0. 05）. Compared to the negative control group, the transfection of miR-411 mimic inhibited the proliferation, migration and invasion of MDA-MB-231 breast cancer cells （P 〈0. 05）. Targetscan showed that miR-411 could bind to GRB2 3 ＇UTR at position 741-747. Compared with the negative control group, GRB2 3 ＇UTR wild-type plasmid and miR-411 co-transfection reduced the fluorescence activity （P 〈 0. 05 ）. Transfection of siGRB2 significantly reduced the expression of GRB2 protein in MDA-MB-231 breast cancer cells （P 〈 0. 05）. Compared to the negative control group, the inhibition of GRB2 expression reduced the proliferation and the number of colony formation of MDA-MB-23

关 键 词：微RNAs/代谢 乳腺肿瘤/代谢 细胞增殖 肿瘤转移 

分 类 号：R737.9[医药卫生—肿瘤]