靶向survivin的反义寡核苷酸联合三苯氧胺治疗乳腺癌的研究  

Experimental Study of Anti-Breast Cancer Cells with Tamoxifen Combined with Antisense Oligonucleotide Targeting Survivin mRNA

作　　者：宋文哲[1] 宋燕[2] 谭岩[3] 方艳秋[3] 

机构地区：[1]徐州医学院附属医院普通外科,江苏徐州221002 [2]吉林大学中日联谊医院基本外科,长春130033 [3]吉林大学第一医院中心研究室,长春130021

出　　处：《中国普外基础与临床杂志》2010年第9期922-926,共5页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的研究联合应用靶向survivin的反义寡核苷酸(ASODN)和三苯氧胺(TAM)对MCF-7乳腺癌细胞的作用。方法将一条20mer的ASODN序列与TAM作用于MCF-7乳腺癌细胞,分别为TAM组(单独应用TAM)、ASODN组(单独应用ASODN)及TAM+ASOND联合组(联合应用TAM+ASODN).运用四甲基偶氮唑盐比色试验(MTT)、流式细胞仪(FCM)、原位杂交以及分光光度法分别检测各组MCF-7细胞的抑制率、细胞周期和凋亡率、survivin mRNA表达和caspase-3活性。结果 TAM+ASOND联合组对MCF-7细胞的抑制率〔(62.26±3.92)%〕明显高于TAM组〔(42.30±6.63)%〕及ASODN组〔(54.77±9.99)%〕,P<0.05.TAM+ASOND联合组的细胞凋亡率为(28.08±4.32)%,明显高于TAM组〔(18.94±4.01)%〕及ASODN组〔(21.12±3.95)%〕,P<0.01.TAM+ASODN联合组对MCF-7细胞的G0/G1期阻滞作用明显强于TAM组和ASODN组(P<0.05,P<0.01);TAM+ASODN联合组survivin mRNA表达阳性率为(13.38±3.45)%,明显低于TAM组〔(39.67±7.42)%〕或ASODN组〔(27.50±5.80)%〕,P<0.01.TAM+ASOND联合组MCF-7细胞的caspase-3活性(0.93±0.13)高于TAM组(0.50±0.09)或ASODN组(0.64±0.08),P<0.01.结论靶向survivin的反义寡核苷酸能够促进MCF-7乳腺癌细胞凋亡,增加其对TAM治疗的敏感性。Objective To study the effects on MCF-7 breast cancer cells with combination of tamoxifen（TAM） and antisense oligonucleotide （ASODN） targeting survivin mRNA. Methods MCF-7 breast cancer cells were treated with a 20 mer ASODN targeting survivin mRNA and TAM,which were divided into three groups：TAM group （treated by TAM only）,ASODN group （by ASODN only）,and TAM＋ASODN combined group （by TAM＋ASODN combination）. The growth inhibition of MCF-7 cells,the changes of cell cycles and apoptotic rate,the positive rate of survivin mRNA expression,and the activity of caspase-3 were tested by MTT,flow cytometry,hybridization in situ,and spectrophotometric method,respectively. Results The rate of growth inhibition of MCF-7 cells in the TAM＋ASODN combined group was （62.26±3.92）%,which was significantly higher than that in the TAM group 〔（42.30±6.63）%〕 or ASODN group 〔（54.77±9.99）%〕,P0.05. The apoptotic rate of MCF-7 cells was （28.08±4.32）% in the TAM＋ASODN combined group,which was significantly higher than that in the TAM group 〔（18.94±4.01）%〕 or ASODN group 〔（21.12±3.95）%〕,P0.01. The effect of arresting MCF-7 cells in G0/G1 phase in the TAM＋ASODN combined group was stronger than that in the TAM or ASODN group （P0.05,P0.01）. The positive rate of survivin mRNA in the TAM＋ASODN combined group was （13.38±3.45）%,which was significantly lower than that in the TAM group 〔（39.67±7.42）%〕 or ASODN group 〔（27.50±5.80）%〕,P0.01. The activity of caspase-3 in the TAM＋ASODN combined group （0.93±0.13） was significantly higher than that in the TAM group （0.50±0.09） or ASODN group （0.64±0.08）,P0.01. Conclusion The ASODN targeting survivin mRNA can promote the apoptosis of MCF-7 breast cancer cells,and make MCF-7 cells more sensitive to tamoxifen.

关 键 词：乳腺癌 三苯氧胺 SURVIVIN 反义寡核苷酸 CASPASE-3 

分 类 号：R737.9[医药卫生—肿瘤]