胆囊癌细胞中转染Survivin和bcl-2反义寡核苷酸后凋亡作用的对比研究  

The comparative study about the apoptosis induced by antisense oligonucleotide targeting Survivin and bcl-2 on gall bladder cancer cell line GBC-SD in vitro

作　　者：冯立民[1] 姜希宏[1] 乌新林[1] 苏永红[1] 徐克森[1] 

机构地区：[1]山东大学齐鲁医院普外科,山东济南250012

出　　处：《中国现代普通外科进展》2005年第2期87-89,共3页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:比较转染Survivin、bcl -2反义寡核苷酸(ASODN)对胆囊癌细胞系 GBC SD的凋亡促进作用。方法:设计并合成特异性靶向 Survivin及 bcl- 2 的 ASODN。胆囊癌细胞分为 5 组:空白对照组、Survivin ASODN转染组、bcl- 2 ASODN转染组、Survivin SODN转染组和bcl -2 SODN转染组,转染24h后,采用逆转录 聚合酶链反应(RT PCR)检测Survivin基因、bcl- 2基因表达的变化,流式细胞仪技术检测细胞凋亡率,四甲基偶氮唑蓝(MTT)法检测 ASODN对细胞的生长抑制率。结果:转染 Survivin及bcl 2 ASODN后,胆囊癌细胞内Survivin基因表达下调 47.8%,bcl- 2 基因表达下调 52.4%;细胞凋亡率分别为(11.4±3. 9)%、(7. 3±3. 2)%,与对照组差异有统计学意义(P< 0. 01)。而转染 SurvivinASODN组及bc1 2ASODN组相比差异也有显著性(P<0.05)。两转染组相对于空白对照组的生长抑制率分别为54.3%、41.6%。结论:Survivin和bcl 2反义寡核苷酸均可有效的抑制Survivin及bc1- 2基因的表达,从而诱导胆囊癌细胞的凋亡,两者相比,Survivin反义寡核苷酸可更为有效地诱导胆囊癌细胞的凋亡。Objective ：To investigate the apoptosis induced by antisense olisonucleotide targeting Survivin and bcl-2 on gallbladder cancer line GBC-SD in vitro.Methods ： ASODN tarseting Survivin and bcl-2 was desisned and constructed. Cultrued cells were divided into 5 groups： control group, Survivin ASODN group, bcl-2 ASODN group, Survivin SODN goup, bcl-2 SODN group. After transfected for 24 h, the cultured ceils were harvested to carry out the next tests. The expression of Survivin and bcl-2 was detected by RT-PCR; Apoptosls index（AI） was examined by flow cytometry; Inhibitory rate（IR）by ASODN was determined by the colorimetrl MTT cell viability and proliferation assay. Results ：The expression of Survivin and bcl-2 were decreased 47.83% and 52.35% resloectively. The AI in Survivin ASODN group and bcl-2 ASODN group was 11.38%±3.91% and 7.26%±3.15% respectively and significantly higher than the control group（P〈0.01）,and that in the Survivin ASCDN group was significantly higher than the bcl-2 ASCDN group,（P〈0.05）.The IR was 54.3% and 41.6% respectively.Conclusion：The expression of Survivin and bcl-2 may be decreased in GBC-SD cells after Survivin and bcl-2 ASODN transfection,and the apcptosis was induced.Comparing the two ASODNs,Survivin ASODN was more effectively.

关 键 词：胆囊肿瘤 基因 Survivin 基因 BCL-2 寡核苷酸类 反义 转染 

分 类 号：R735.8[医药卫生—肿瘤]