小干扰RNA沉默Y盒结合蛋白-1基因对肺腺癌A549细胞株表皮生长因子受体表达的影响  

Expression of epidermal growth factor receptor in lung adenocarcinoma after silencing Y box-bind- ing protein-1 gene by small interfering

作　　者：赵士磊[1] 赵玉达[1] 尧然[1] 李锦绣[1] 张嘉宁[2] 顾春东[1] 

机构地区：[1]大连医科大学附属第一医院胸外科肺癌诊疗基地,116011 [2]大连医科大学基础医学院糖生化重点实验室

出　　处：《中华实验外科杂志》2013年第8期1595-1597,共3页Chinese Journal of Experimental Surgery

基　　金：基国家自然科学基金资助项目（81173453）;大连市科技计划重大项目（2012E15SF141）

摘　　要：目的观察Y盒结合蛋白-1（YB-1）小干扰RNA（siRNA）对肺腺癌A549细胞株表皮生长因子受体（EGFR）的影响。方法针对YB-1mRNA设计特异性siRNA及ControlsiRNA序列转染至A549细胞株，采用逆转录-聚合酶链反应（RT-PCR）和Westernblot检测空白组、单纯Lipofectamine2000处理组、ControlsiRNA组及YB-1siRNA组细胞中YB-1和EGFR在mRNA和蛋白水平的表达；通过噻唑蓝（MTT）、侵袭实验观察各组细胞的生物学行为变化。结果RT-PCR和Westernblot结果显示实验组YB-1与EGFR的mRNA和蛋白表达降低（分别为0．3820±0．0094、0．4690±0．0032和0．1820±0．0073、0．4210±0．0049），与对照组比较差异有统计学意义（P〈0．05）。MTT检测转染72h后YB-1 siRNA组吸光度值为1．34±0．11，同时转染48h后YB-1siRNA组侵袭能力较对照组明显降低（P〈0．05）。结论YB-1siRNA能抑制肺腺癌A549细胞株中EGFR的表达，影响肿瘤增殖和侵袭能力。Objective To investigate the expression of epidermal growth factor receptor （EGFR） using the small interfering RNA （siRNA） targeting Y box-binding protein-1 （YB-1） gene in lung adeno- carcinoma cell line A549. Methods Specific siRNA and a negative control sequence in accordance with YB-1 mRNA from GeneBank were designed and then transfected into A549 cells separately. In blank con- trol group, Lipofectamine 2000 treatment group, negative control siRNA group and YB-1 siRNA group, re- verse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting were applied to detect the changes of YB-1 and EGFR mRNA and protein expression. Cell proliferation was examined by using MTT assay. The ability of invasion was tested by using cell invasion assay. Results RT-PCR results showed that the expression of YB-1 and EGFR mRNA （0. 3820 ±0. 0094 and 0. 4690 ±0. 0032） in experimental group was decreased significantly as compared with other groups （P 〈 0. 01 for all） , and Western blotting results also showed the expression of YB-1 and EGFR proteins （ 0. 3820 ± 0. 0094 and 0. 4690 ± 0. 0032 ） in ex- perimental group was significantly lower than other groups （ P 〈 0. 05 for all）. After transfection the prolif- eration and invasive ability of A549 cells were significantly reduced in experimental group as compared with other control groups. Conclusion Silencing the expression of YB-1 gene by using siRNA can inhibit the expression of EGFR in lung adenocareinoma A549 cell line and thus adjust proliferation and invasion of tumor.

关 键 词：Y盒结合蛋白-1基因 小干扰RNA 表皮生长因子受体 肺腺癌 

分 类 号：R734.2[医药卫生—肿瘤]