脂质体携带反义寡核苷酸对A375细胞Survivin表达的影响  

Effects of Antisense Oligonucleotide Carried by the Liposome-encapsulation on Expression of Survivin mRNA in A375 Cell

作　　者：王梅[1] 谭升顺[1] 王冰[2] 

机构地区：[1]西安交通大学医学院第二附属医院皮肤科,陕西西安710004 [2]西安交通大学医学院病理系,陕西西安710061

出　　处：《中国皮肤性病学杂志》2007年第9期521-523,共3页The Chinese Journal of Dermatovenereology

基　　金：陕西省科技攻关基金资助项目(No.2003K10-G10)

摘　　要：目的研究脂质体携带反义寡核苷酸转染对A375细胞Survivin mRNA及蛋白表达的影响。方法通过反转录聚合酶链反应(RT-PCR)和Western免疫印迹法对反义寡核苷酸转染后A375细胞Survivin mRNA和蛋白表达变化进行定性和半定量分析。结果RT-PCR结果经光密度扫描半定量分析:对照组为0.9994,反义寡核苷酸处理组为0.4761,较对照组降低了52.36%。Western免疫印迹法证实在A375细胞中存在分子量16.5KD的特异性条带,与Survivin分子量相符,反义寡核苷酸处理组细胞在NC膜上特异性条带明显弱于对照组细胞。结论证实反义寡核苷酸通过降解mRNA,阻断基因翻译成蛋白质的特异性反义活动机制。Objective To observe effects of survivin antisense oligonucleotide by the liposome-encapsulation on expression of survivin mRNA and proteins in A375 cells. Methods After antisense oligonucleotide transfected, survivin gene and proteins expression alteration were qualitation and semi-quantitive analyzed by use of RT-PCR and Western blot methods. Results By use of RT-PCR methods, survivin gene expression alteration was analyzed. The quantity PCR product was determined by scanning of fluorescent intensity, in comparison with GAPDH, survivin mRNA semi-quantitive results were as follows： control group： 0. 9994, ASODN treatment group：0.4761 ,the percentages of inhibition vs untreated controls was 52.36%. Expression of survivin in malignant melanoma cell line A375 was confirmed by Western blot, demonstrating a - 16.5KD survivin band. Survivin proteins expression alteration was analyzed. It was clear that specific band on NC membrane of transfected cell was obviously weaken compared with that in control cell by immunoblotting. Conclusion Results demonstrated antisense oligonucleotide specific mechanisms of action,by inhibiting transcription and degradating targeting mRNA, ASODN treatment can downregulate the survivin mRNA level, and further block the reading of that genetic information, reduce the survivin protein expression.

关 键 词：SURVIVIN 恶性黑色素瘤 反义寡核苷酸 

分 类 号：R739.5[医药卫生—肿瘤]