【摘要】 目的研究小干扰RNA(siRNA)对类表皮生长因子域7(egfl7)基因表达的抑制作用。方法利用Ambion公司设计合成的以egfl7为靶标的siRNA,通过脂质体将siRNA转入人脐静脉内皮细胞株,以未转染siRNA细胞和转染无关siRNA细胞为对照,利用MTS[3-(4,5-dimethylthiaz-2-yl)- 5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,inner salt]法检测siRNA对细胞存活率的影响,同时检测乳酸脱氢酶和三磷酸腺苷,观察siRNA对细胞生长的影响,RT-PCB法检测egfl7 mRNA水平的改变,Western blot检测egfl7蛋白表达的变化。结果siRNA各组与对照组细胞存活率差异均有统计学意义(P<0．05),乳酸脱氢酶及三磷酸腺苷释放量差异也存在统计学意义(P< 0．01)。转染24 h后,siRNA组egfl7 mBNA与对照组相比差异有统计学意义(P<0．01),同时egfl7蛋白表达也被明显抑制(P<0．01)。其中siBNA1的抑制效率最高。结论体外转录合成的siRNA可抑制人脐静脉内皮细胞egfl7基因的表达。更多还原

【Abstract】 Objective To evaluate the effects of small interference RNA (siRNA) on epidermal growth factor-like domain 7 (egfl7) gene expression in human endothelial cell line HUVEC. Methods siRNA targeting egfl7 (siRNA1, siRNA2, siRNA3 and siRNA4) was constructed through online design of Ambion company and transfected into human endothelial cell line HUVEC with lipofectamine. The nontransfected cells and cells treated with control siRNA were taken as controls. At 24, 48 and 72 hours post various interventions, cell viability was determined by MTS method as well as LDH and ATP releasing tests. egfl7 expressions at protein and mRNA levels were detected by Western blot and RT-PCR respectively. Results Cell survival rate, LDH and ATP release were significantly reduced in siRNA treated cells compared to control cells (P<0. 05). Similarly, egfl7 expression at protein and mRNA levels was also significantly reduced in siRNA treated cells (P<0.01) , especially in siRNA1 treated cells. Conclusion siRNA inhibited egfl7 gene expression and cell survival in HUVEC.更多还原