微小RNA-335靶向Sp1对乳腺癌细胞株MCF-7增殖的调控作用  

MicroRNA-335 regulates proliferation of breast cancer cells MCF-7 by targeting Sp1

作　　者：杨浚讽 李文环[2] 李海[2] 钟源[2] 江学庆[2] 孙圣荣[1] 

机构地区：[1]武汉大学人民医院乳腺甲状腺外科,430060 [2]武汉市中心医院甲状腺乳腺外科

出　　处：《中华实验外科杂志》2015年第11期2688-2690,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察微小RNA （miR）-335靶向Sp1对乳腺癌细胞株MCF-7增殖的影响.方法 选取MCF-7为研究对象,采用荧光素酶报告基因实验验证miR-335对Sp1的靶向作用;采用反转录-聚合酶链反应（RT-PCR）检测细胞中miR-335及Sp1 mRNA表达;采用Western blot检测细胞中Sp1蛋白表达;采用噻唑蓝（MTT）法检测MCF-7增殖水平.结果 与正常乳腺上皮细胞株HBL-100比较,MCF-7中miR-335表达显著降低（0.93 ±0.11比2.94±0.13）,Sp1蛋白表达显著增加（0.89±0.08比0.42±0.05,P＜0.01）.miR-335可与Sp1基因3＇-非翻译区（UTR）特异性结合.转染miR-335 mimic可使MCF-7中miR-335表达显著提高,Sp1蛋白表达显著降低,细胞增殖受到抑制;转染miR-335 mimic＋ Sp1可使Sp1蛋白表达显著提高,同时部分逆转miR-335对MCF-7细胞增殖的抑制作用.结论 miR-335可通过靶向Sp1抑制乳腺癌细胞株MCF-7增殖.Objective To evaluate the regulatory effect of microRNA （miR）-335 targeting Sp1 on the proliferation of breast cancer cells MCF-7.Methods By using MCF-7 cells, luciferase reporter gene assay was used to detect the targeting effect of miR-335 to Sp1.Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to detect the mRNA expression of miR-335 and Sp1.Western blotting was used to detect the protein expression of Sp1.MTT method was used to detect the proliferation of MCF-7 cells.Results Compared with normal breast epithelium HBL-100, the miR-335 expression of MCF-7 cells was significantly reduced [（0.93 ±0.11） vs.（2.94 ±0.13）], and the Sp1 protein expression in MCF-7 cells was significantly increased [（0.89 ± 0.08） vs.（0.42 ± 0.05）, P 〈 0.01].miR-335 was specifically bound with Sp1 3＇-untranslated region （UTR）.After transfection of miR-335 mimic into MCF-7 cells, the miR-335 expression significantly increased, the Sp1 protein expression significantly reduced, and cell proliferation was significantly restrained.After transfection of miR-335 mimic ＋ Sp1 into MCF-7 cells, the Sp1 expression significantly increased, and partly reversed the inhibitory effect of miR-335 on cell proliferation.Conclusion miR-335 can inhibit proliferation of MCF-7 cells through targeting Sp1.

关 键 词：微小RNA-335 SP1 乳腺癌 细胞增殖 MicroRNA-335 SP1 

分 类 号：R155.3[医药卫生—营养与食品卫生学]