摘要
目的:观察iASPP基因干扰RNA转染乳腺癌细胞MCF-7后的干扰效果及细胞凋亡变化。方法:设计特异性siRNA序列,将序列克隆至PGCsilencerTM H1/Neo/GFP质粒中,用脂质体将重组子转染至MCF-7细胞中,用RT-PCR方法检测i ASPP的表达,蛋白质印迹法检测蛋白表达的变化,流式细胞仪检测细胞凋亡的情况。结果:iASPP干扰质粒转染MCF-7细胞后,iASPP的mRNA表达和蛋白表达减少40%～50%;p53蛋白相对表达量由转染阴性质粒的0.37增加到转染干扰质粒的0.64;细胞凋亡率由原来的17.8%和16.2%分别增加到53.5%和51.3%。结论:抑制内源性i ASPP能有效地恢复乳腺癌细胞MCF-7中p53的抑癌功能,为乳腺癌的治疗提供新的思路。
OBJECTIVE： To observe the effect of the iASPP RNAi vector transfection on the apoptosis of breat cancer cell MCF7. METHODS： The specific siRNA sequence was designed according to the genebank. The se quence was cloned into PGCsilencerTM H1/Neo/GFP. The recombinant plasmid was transfected into MCF-7 by Lipofeetamine^TM 2000. The iASPP expression was analyzed by RT-PCR and Western blot. The cell apoptosis was detected by FCM. RESULTS： The iASPP expression in MCF-7 cell lines was descended from 40% to 50% after the RNAi transfection. The expression of p53 protein increased from 0.37 to 0.64, and the apoptosis rate increased from 17.8% and 16.2% to 53.5% and 51.3%. CONCLUSION：The inhibition of endogenous iASPP may resume the apoptosis activity of p53 in MCF-7.
出处
《中华肿瘤防治杂志》
CAS
2008年第21期1605-1607,1635,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(30428013,30470675)
关键词
乳腺肿瘤
基因
肿瘤抑制
RNA干扰
细胞凋亡
breast neoplasms
genes, tumor suppressor
RNA interference
cell apoptosis
