mirRNA―21逆向调控REK促进乳腺癌细胞侵袭转移的研究.docVIP

microRNA-21逆向调控RECK促进乳腺癌细胞侵袭转移的研究 】目的研究microRNA-21参与乳腺癌 侵袭转移的调控机制。方法采用实时定量反转录- 聚合酶反应(RT-PCR)方法检测MDA-435、MDA-231 和MCF-7乳腺癌细胞株microRNA-21的表达；采用 trans well方法测定上述三株乳腺癌细胞体外侵袭转移 能力；人工合成microRNA-21干扰序列，转染 MDA-231、MDA-435乳腺癌细胞，观察对细胞侵 袭能力的影响；应用Western blot检测逆转诱导蛋白 (RECK)的表达；采用焚光报告基因实验检测RECK 与 microRNA-21 的结合情况。结果 MCF-7、MDA-231 及MDA-435细胞株中均具有microRNA-21高表 达，且表达量 MDA-435MDA-231 MCF-7,其 中，具有高侵袭特性的MDA-435细胞具有最高水 平 microRNA-21 的表达为(4.51 ±0.71)，MDA-231 细胞具有中等水平的microRNA-21表达为(2.67土 0.27)，MCF-7细胞表达microRNA-21水平相对较低 为(1.23±0.11)，比较差异有统计学意义(P 关键词】乳腺癌；侵袭；microRNA-21;伴Kazal 域的富含半胱氨酸的逆转诱导蛋白；RNA干扰 DOI： 10.14163/j.cnki.ll-5547/r.2016.16.195 Abstract】 Objective To research regulation mechanism of microRNA-21 for breast cancer invasion and metastasis. Methods Real-time quantification polymerase chain reaction (RT-PCR) was applied to detect miR-21 expression in MDA-435，MDA-231 and MCF-7 breast cancer strains. In vitro invasion and metastasis of these three strains were detected by transwell method. Transfection on MDA-231 and MDA-435 cell was made by synthetic microRNA 21 interference sequence to observe the influence on cell invasion. Reversion-inducing cysteine-richprotein with Kazalmotifs (RECK) protein expression was detected by Western blot， and combination of RECK and microRNA-21 was detected by luciferase reporter gene test. Results All MCF-7， MDA-231 and MDA-435 strains had high expressed microRNA-21， and their expression volume were shown as MDA-435MDA-231 MCF-7. MDA-435 with high invasion feature showed the highest microRNA-21 expression level as (4.51 ±0.71followed by microRNA-21 expression in MDA-231 as (2.67 ±0.27) and in MCF-7 as (1.23 ±0.11)， and their difference had statistical significance (P Key words】 Breast cancer； Invasion； microRNA-21 ； Reversion-inducing cysteine-richprotein with Kazalmotifs ； RNA interference microRNAs是新近识别的人体内一类内源性非编 码小分子核糖核酸(RNA)，通过与靶基因互补结 合，在转录后水平调控靶蛋白表达，从而参与生 物体各项生物学活动。前期研宄显示［1］， microRNA-21在乳腺癌细胞中高表达并与阿霉素耐药 有关，并有研宄提示［2］，microRNA-21高表达与 乳腺癌