摘要
目的应用RNA干扰技术靶向阻断survivin基因表达,探讨其蛋白表达水平及对细胞增殖抑制作用。方法将siRNA真核表达载体Pgenesil-sur通过脂质体介导转染腺样囊性癌细胞,采用免疫组化法检测survivin基因蛋白表达;并用图象分析仪分析蛋白表达强度及表达抑制率。MTT法检测对腺样囊性癌细胞增殖的抑制作用。结果空白对照组、空质粒载体组、RNA干扰组survivin蛋白表达强度分别为:2.52±0.10,2.48±0.11,0.53±0.10。经统计学分析表明转染Pgenesil-sur真核表达载体后,survivin基因蛋白表达水平明显减低(P<0.05),其抑制率达到79%;转染空质粒载体与空白对照组差异无统计学意义(P>0.05);MTT法结果表明转染Pgenesil-sur真核表达载体后细胞增殖受到显著抑制,抑制率为38.80%(P<0.05);转染空质粒载体细胞增殖未受到抑制(P>0.05)。结论靶向survivin siRNA能抑制体外培养的腺样囊性癌细胞survivin基因蛋白表达与细胞增殖作用。
Objective To investigate the effect of siRNA in suppression survivin expression and inhibition effect of cell proliferation in adenoid cystic carcinoma （ACC）. Methods SiRNA eukaryotic expression vector Pgenesil/sur was used to transfect the cells of adenoid cystic carcinoma（ACC） through liposome and the expression of survivin was tested by immunohistochemical. The survivin expression and inhibition rate were analysed by the image instrument. The effect of siRNA in suppressing the proliferation of adenoid cystic careinoma（ACC） cells was detected by MTT assay, Results The survivin expression strength of the negative control group,negative plasmid vector group,RNA interference group respec- tively was：2.52±0. 10,2.48±0. 11 and 0. 53±0.10. Analysed by statistics indicated that after using siRNA eukaryotic expression vector Pgenesil/sur transfected the cell of adenoid cystic earcinoma（ACC）, the level of the survivin expression decreased obviously（P〈0.05）. The rate of inhibition reach to 79%. The difference between transfected negative plasmid vector and control group was insignificant （P〈0.05）. The results of MTT assay indicated that after transfected eukaryotic expression vector Pgenesil/sur,cell proliferation was inhibited obviously,inhibition rate was 38.80% （P〈0.05）. Transfected negative plasmid vector the cell proliferation was unchanged（P〉0. 05）. Conclusion The siRNA eukaryotic expression vector target against survivin constructed in the study can block the expression of survivin and cell proliferation in adenoid cystic carcinoma（ACC） ceil effectively and specifically.
出处
《现代检验医学杂志》
CAS
2008年第5期21-23,共3页
Journal of Modern Laboratory Medicine
