摘要
目的探讨慢病毒载体介导survivin-siRNA对人肺腺癌裸鼠移植瘤的体内抑瘤活性。方法参考siRNA的设计策略,构建表达survivin-siRNA的慢病毒载体;于各BALB/C裸鼠右侧腋窝皮下接种人肺腺癌A549细胞悬液,构建人肺腺癌A549细胞裸鼠移植瘤模型,肿瘤组织局部注射survivin-siRNA的慢病毒载体,观察肿瘤的体积及随时间变化的生长曲线;通过碘化丙啶（propidiumiodide,PI）染色检测细胞凋亡情况;流式细胞术检测肿瘤细胞周期变化。结果慢病毒载体介导survivin-siRNA对裸鼠人肺腺癌A549的抑瘤率为46.07%;经过PI和Annexin-V染色区分凋亡早晚期的细胞为30%-35%,荧光显微镜下观察肿瘤细胞凋亡;G1期细胞比例明显增加,S期细胞比例则明显减少。结论慢病毒载体介导的siRNA能有效抑制裸鼠移植人肺腺癌A549的survivin基因的表达,有效激发细胞凋亡。
Objective To explore tumor inhibition of human lung adenocarcinoma nude mice by the lentiviral vectors mediate siRNA for survivin gene in vivo. Methods Referencing siRNA design strategy to construct expression of survivin-siRNA lentiviral vector;we in all BALB/C nude mice inoculated human lung adenocarcinoma A549 cell suspension into subcutaneously of the right armpit.Human lung adenocarcinoma A549 nude mice model was constructed,and the lentiviral vectors were local injected in tumor tissue.Then the curve about the changing of tumor volume in different time were observed.The histogram of tumor cell cycle and apoptosis were examined by PI and FCM,respectively.Results The inhibition rate of human lung adenocarcinoma A549 tumor of lentiviral vector mediated survivin-siRNA in nude mice was 46.07%;30%~35%by the PI and Annexin-V staining to differentiate between early late apoptotic cells and dead cells. Apoptosis of tumor cells were observed under fluorescence microscopy.G1 phase cells significantly increased,S phase cells were decreased significantly. Conclusion The lentiviral vectors mediated siRNA for survivin can significantly inhibit survivin gene express in vivo and markedly induce the apoptosis of A549 tumor.
出处
《中国医药生物技术》
CSCD
2010年第2期117-121,共5页
Chinese Medicinal Biotechnology
基金
聊城市科技攻关计划资助项目(20070248)
