小干扰RNA抑制胚胎腭突间充质细胞7-脱氢胆固醇还原酶基因的表达  

Inhibitory effect of small interfering RNA on expression of 7-dehydrocholestererol reductase gene inembryo palatal mesenchymal cells

作　　者：庄翠竹[1] 马秀兴[1] 肖文林[1] 张春阳[1] 王双义[1] 

机构地区：[1]青岛大学医学院黄岛院区口腔科,266003

出　　处：《中华实验外科杂志》2012年第11期2283-2285,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81070817）;山东省自然科学基金资助项目（ZR2010HM054）

摘　　要：目的观察小鼠胚胎腭突间充质（EPM）细胞中7-脱氢胆固醇还原酶基因（Dhcr7）的表达，筛选出针对EPM细胞Dhcr7的最有效小干扰RNA（siRNA）序列。观察RNA干扰（RNAi）沉默Dhcr7的表达后，对EPM细胞增殖的影响。方法化学合成3条针对Dhcr7mRNA的siRNA，阳性脂质体介导转染EPM细胞。荧光显微镜观察转染效率，Westernblot法检测Dhcr7蛋白表达变化，噻唑蓝（MTT）检测沉默后增殖率的变化。结果siRNA转染效率为72．6％。3条siRNA对EPM细胞Dhcr7基因表达都有抑制作用，siRNA-3抑制作用最明显，达60．2％，与阴性对照组比较差异有统计学意义（P〈0．05）。Dhcr7沉默后EPM细胞增殖抑制率为56．4％，与阴性对照组比较差异有统计学意义（P〈0．01）。结论筛选出针对EPM细胞Dhcr7最有效的siRNA序列。siRNA可抑制EPM细胞Dhcr7基因表达，Dhcr7表达降低可抑制EPM细胞增殖。Objective To study the expression of 7-dehydrocholestererol reductase gene （Dhcr7） in the embryo palatal mesenchymal （ EPM ） cells, to screen out the most effective sequence of Dhcr7 siRNA, and investigate the effects of siRNA-mediated silencing of the Dhcr7 gene on the expression and proliferation of EPM ceils. Methods Three small fragments of siRNA chemically synthesized against Dhcr7 mRNA were transfected into EPM cells by positive liposomal transfection. The transfection efficiency was observed by fluorescence microscope, the protein expression of Dhcr7 was examined by Western blot- ting, and the changes in proliferation were evaluated by methyl thiazol tetrazolium （MTT） assay. Results The transfection efficiency was about 72. 6%. The expression levels of Dhcr7 were inhibited by the three Dhcr7 siRNAs, but siRNA-3 was the most effective one, and the protein expression of Dhcr7 was reduced by 60. 2% , which was significantly lower than the control group （ P 〈 0. 05 ）. The proliferation rate of EPM ceils transfected with Dhcr7 siRNA was 56. 4% , which was significantly lower than the control group （ P 〈 0. 01 ）. Conclusion This study screened out the most effective sequence of Dhcr7 siRNA, which Could inhibit the Dhcr7 expression and proliferation of EPM cells.

关 键 词：7-脱氢胆固醇还原酶 腭突间充质细胞 RNA干扰 

分 类 号：R782.2[医药卫生—口腔医学]