摘要
目的探讨"三阴"乳腺癌细胞中沉默survivin基因对抑癌基因caspase3和caspase7的影响。方法针对survivin基因设计干扰片段并与真核表达载体pGCsilencerTMU6/Neo/GFP进行连接,之后转染"三阴"乳腺癌细胞MDA-MB-231;通过RT-PCR鉴定survivin基因mRNA水平沉默效果;用RT-PCR检测沉默survivin基因对caspase3和caspase7表达及细胞凋亡的影响。结果 survivin基因沉默可引起MDA-MB-231细胞凋亡;caspase3和caspase7表达升高,灰度分析结果显示差异具有统计学意义(P=0.008)。结论 (1)RNAi技术沉默MDA-MB-231细胞中survivin基因,达到了对其表达的抑制效果;(2)survivin基因沉默可上调caspase3和caspase7的表达;(3)survivin基因沉默可引起细胞凋亡;(4)survivin基因可作为"三阴"乳腺癌基因治疗的新靶点。
Objective To explore the effects of gene silencing of survivin on caspase3 and caspase7 in breast cancer cell line MDA-MB-231.Methods shRNA fragment was designed and inserted to eukaryotic expression vector pGCsilencer TMU6/Neo/GFP,and was transfected into MDA-MB-231 cells.The expression of surviving,caspase3 and caspase7 were detected by RT-PCR and western blot.Results Surviving gene silencing in MDA-MB-231 could up regulate the apoptosis ability of cell; survivin silence could increase the expression of caspase3 and caspase7 （P =0.008）.Conclusions （1） Designed RNAi fragment and transfected it into MDA-MB-231 cell,it could silence surviving gene; （2）Gene silence of survivin could increase expression of caspase3 and caspase7 ; （3）Gene silence of survivin could increase the apoptosis ability of cell; （4） Gene silence of survivin could been new target gene in HR-and HER-breast cancer therapy.
出处
《中华临床医师杂志（电子版）》
CAS
2013年第1期103-105,共3页
Chinese Journal of Clinicians(Electronic Edition)
基金
吉林省教育厅基金(吉教科合字200687)
