靶向survivin基因反义寡核苷酸对MCF-7人乳腺癌细胞系凋亡及药物敏感性等生物学行为的影响  

Effect of Antisense Oligonucleotide Targeting Survivin Gene on Biological Behaviors of Apoptosis and Drug Sensitivity of MCF-7 Human Breast Cancer Cell Line

作　　者：高砚春[1] 陈竞[2] 李金穗[1] 

机构地区：[1]川北医学院附属医院普外三科,四川南充637007 [2]川北医学院基础医学院生物化学教研室,四川南充637007

出　　处：《医学综述》2016年第14期2865-2868,F0003,共5页Medical Recapitulate

摘　　要：目的探讨靶向survivin基因反义寡核苷酸(as ON)对MCF-7人乳腺癌细胞系凋亡及药物敏感性等生物学行为的影响。方法 MCF-7人乳腺癌细胞株分为正常组、脂质体组、正义组和反义组,正常组常规培养,并用Opti-MEM培养液代替转染液进行转染;脂质体组采用Lipofectamine 2000-Opti-MEM代替转染液进行转染;正义组采用survivin正义寡核苷酸进行转染;反义组采用survivin基因as ON转染。采用反转录聚合酶链反应检测各组survivin mRNA表达水平,四甲基偶氮唑盐法检测细胞抑制率;分别用紫杉醇处理MCF-7细胞株,并检测紫杉醇处理后survivin mRNA表达水平和细胞抑制率。结果 as ON转染后,MCF-7细胞中survivin mRNA表达水平显著降低,其中反义组survivin mRNA表达水平显著低于正常组、脂质体组和正义组(0.34±0.10比0.63±0.10、0.74±0.20、0.73±0.10),细胞抑制率显著高于正常组、脂质体组和正义组[(51.32±7.42)%比(3.72±0.10)%、(4.12±0.10)%、(4.53±0.10)%],差异有统计学意义(P<0.05)。转染后反义组细胞周期被阻滞于G2/M期;其中反义组G2/M期细胞比率显著高于正常组、脂质体组和正义组[(34.9±2.7)%比(21.5±2.8)%、(19.4±2.6)%、(18.8±1.9)%],反义组细胞凋亡率亦显著高于正常组、脂质体组和正义组[(16.3±1.2)%比(5.1±0.4)%、(5.3±0.5)%、(5.7±0.5)%],差异有统计学意义(P<0.05)。各组经紫杉醇处理后survivin mRNA表达水平和细胞抑制率均显著增加,与不加药组比较差异有统计学意义(P<0.05);其中反义组survivin mRNA表达水平显著低于正常组、脂质体组和正义组(0.62±0.15比1.37±0.33、1.47±0.33、1.33±0.26),细胞抑制率显著高于正常组、脂质体组和正义组[(84.75±9.62)%比(36.44±3.73)%、(41.45±5.36)%、(40.82±4.93)%],差异有统计学意义(P<0.05)。结论 survivin基因与MCF-7细胞增殖、抗凋亡、化疗耐药等生物学行为有关,通过as ON下调survivin基因的表达水平,能够促进肿瘤细胞�Objective To explore the effect of antisense oligonucleotide targeting survivin gene on bio- logical behaviors of apoptosis and drug sensitivity of of MCF-7 human breast cancer cell line. Methods MCF-7 human breast cancer cell lines were divided into normal group, liposome group, sense group and anti- sense group：the normal group was cultured in Opti-MEM medium instead of the transfection solution;lipo- feetamine 2000-Opti-MEM was used in the liposome group, and the sense group was transfected with survivin oligonucleotide transfection, the anti-sense group was transfected with survivin antisense oligonueleotide （ asON）. The expression level of survivin mRNA was detected by RT-PCR,and the inhibition rate was detec- ted by MIT assay,MCF-7 eel1 lines were treated with taxol,and the expression level of survivin mRNA and cell inhibition rate were detected. Results After asON transfeetion, the expression level of mRNA survivin in MCF-7 cells was significantly decreased, and the expression level of survivin mRNA in the antisense group （0. 34 ±0.10） was significantly lower than that in the normal group（0. 63 ±0. 10） ,liposome group（0.74 ± 0. 20） ,sense group（0. 73 ±0. 10）（P〈0. 05） ,and the inhibition rate（51.32 ±7.42）% was significantly higher than that in normal group（3.72 ±0. 10）% ,liposome group（4. 12 ±0. 10）% ,sense group（4. 53 ± 0. 10）% （P 〈 0.05 ）. After transfeetion, the cell cycle was blocked at G2/M phase, and the G2/M phase cells rate in the anti-sense group （ 34. 9 ± 2.7 ） % was significantly higher than those in the normal group （ 21.5 ± 2.8 ） %, liposome group （ 19.4 ± 2. 6 ） %, and sense group （ 18.8 ± 1.9 ） % （ P 〈 0. 05 ）, and the apoptosis rate（ 16. 3 ± 1.2 ）% was significantly higher than that in the normal group （5.1 ± 0.4 ）% , lipo- some group （5.3 ± O. 5 ） %, and sense group （ 5.7 ± 0. 5 ） % （ P 〈 O. 05 ）. After treatment with taxol, survivin mRNA expression level and cell inhibition rate we

关 键 词：乳腺癌 反义寡核苷酸 细胞凋亡 耐药 凋亡抑制蛋白 

分 类 号：R737.9[医药卫生—肿瘤]