摘要
目的观察Hippo信号通路下游关键因子Yes相关蛋白1（YAP1）对胶质瘤细胞株U87MG细胞增殖周期的影响。
方法使用YAP1特异性小干扰RNA（siRNA）敲低U87MG胶质瘤细胞株YAP的表达，实时定量反转录聚合酶链反应（RT-qPCR）鉴定U87MG胶质瘤细胞中YAP1是否已被敲低；采用RT-qPCR方法分别检测干扰24、48 h后YAP1、细胞核增殖抗原（Ki-67）、c-Myc、血管内皮生长因子（VEGF）、细胞周期蛋白（Cyclin） D1、Cyclin E和p73 mRNA的表达变化。
结果3对siRNA寡核苷酸序列对YAP1都有干扰，第1对siRNA寡核苷酸序列对YAP1有较好的干扰效果，干扰效率可达90%以上，P＝0.044；以1.0为参照，结果显示，YAP1（0.213），Ki-67（0.365），c-Myc（0.618），VEGF（0.534），Cyclin D1（0.540），Cyclin E（0.494），p73（0.314）在24 h出现不同程度下降，在48 h基因的转录水平明显提高；同时可以发现，随着YAP1（0.810）干扰的恢复，Ki-67（1.051），c-Myc（1.498），VEGF（1.057），Cyclin D1（1.598），Cyelin E（0.897），p73（1.110）的活性明显增高，P＝0.007。
结论YAP1调控胶质瘤细胞株U87MG细胞增殖周期，干扰YAP1蛋白后，能阻止细胞增殖于细胞任一周期。
Objective To observe the effect of Hippo Yes associated protein 1 （YAP1） on the proliferation cycle of human glioma cell line U87MG in the downstream of YAP! signaling pathway. Methods YAP specific small interfering RNA （siRNA） was used to knock down the YAP expression in U87MG cells, which was identified by real - time quantitative reverse transcriptase - polymerase chain re- action （RT- qPCR）. Using RT- qPCR method, the expression of YAP1, proliferation cell nuclear anti- gen （Ki -67）, c -Myc, vascular endothelial growth factor （VEGF）, Cyclin D1, Cyclin E and p73 mRNA was detected were used to detect at 24 and 48 h after the interference. Results Three on the siRNA sequence of YAP1 oligonucleotide interference, the first of siRNA oligonucleotide sequence on the YAP1 has a better interference effect, the interference efficiency of up to 90%, P =0. 044. In 1 as a reference, results showed that YAP1 （0. 213）, Ki -67 （0. 365）, c - Myc （0. 618）, VEGF （0. 534）, Cyclin D1 （0. 540）, Cyclin E （0. 494）, p73 （0. 314） in 24 hours decreased to varying degrees, in 48 hours, the level of gene transcription is obviously increased; at the same time that, along with the YAP1 （0. 810） interfereneerecovery, Ki -67 （1.051）, c - Mye （1.498）, VEGF （1.057）, Cyelin D1 （1.598）, Cyelin E （0. 897）, p73 （1. 110） were significantly increased, P=0. 007. Conclusion YAP1 regulates the proliferation cycle of U87MG cells, and after the interference of YAP1 protein, cell proliferation can be arrested at any cell cycle.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第2期243-245,共3页
Chinese Journal of Experimental Surgery
基金
深圳市知识创新计划（JCYJ20140414170821293、JCYJ2013401113155027、JCYJ20130401113503369、JCYJ20150330102401088）
深圳市龙岗区科技计划医疗卫生（扶持类）项目（YLWS20150526165408484）
作者简介
通信作者：王晓彬，Email：evelinew@163．com；
丁建军，Email：dingdang15@163．com
