靶向信号转导和转录激活因子3的小干扰RNA对体内外乳腺癌细胞生长的抑制作用  

Growth inhibition of breast cancer cells in vitro and in vivo by siRNA targeting signal transducers and activators of transcription 3

作　　者：杨增[1] 严丽[1] 张峰[1] 闫庆辉[1] 宋伟庆[1] 王凤安[1] 蔡建辉[1] 

机构地区：[1]河北医科大学第二医院外科,石家庄050000

出　　处：《中华肿瘤杂志》2010年第11期819-824,共6页Chinese Journal of Oncology

摘　　要：目的 观察沉默信号转导和转录激活因子3（STAT3）基因对乳腺癌细胞MCF7体内外生长的抑制作用,探讨STAT3基因作为乳腺癌治疗靶点的可行性和有效性.方法 应用RNA干扰技术抑制MCF7细胞STAT3基因的表达,半定量逆转录聚合酶链反应（RT-PCR）和Western blot法检测STAT3 mRNA及蛋白的表达,流式细胞仪检测MCF7细胞的凋亡率,四甲基偶氮唑蓝（MTT）法检测MCF7细胞的增殖情况.在裸鼠皮下移植转染了STAT3 siRNA的MCF7细胞,观察其成瘤性.半定量RT-PCR和Western blot法检测成瘤标本的STAT3 mRNA和蛋白的表达.结果 转染48 h后,STAT3 siRNA转染组、非特异siRNA转染组和空白对照组的STAT3 mRNA表达量分别为0.327±0.020、1.035±0.050和1.093±0.018;转染72 h后,3组STAT3蛋白表达量分别为0.153±0.006、1.320±0.033和1.374±0.022,STAT3 siRNA转染组的STAT3 mRNA和蛋白表达水平,较非特异siRNA转染组和空白对照组均明显降低（P＜0.05）.MTT实验结果显示,STAT3 siRNA转染组和非特异siRNA转染组的细胞生长抑制率分别为（44.00±5.10）%和（16.10±1.05）%,STAT3 siRNA转染组的细胞增殖能力明显下降（P＜0.05）.流式细胞仪分析显示,STAT3 siRNA转染组、非特异siRNA转染组和空白对照组的细胞凋亡率分别为（14.79±0.22）%、（8.45±0.43）%和（7.06±0.71）%,STAT3 siRNA转染组明显高于非特异siRNA转染组和空白对照组（P＜0.05）.STAT3siRNA转染组、非特异siRNA转染组和空白对照组在裸鼠体内最终的成瘤体积分别为（41.15±12.17）mm3、（101.36±21.90）mm3和（118.45±24.68）mm3,移植瘤重量分别为（21.4±10.6）mg、（57.2±21.9）mg和（88.6±12.2）mg,STAT3 siRNA转染组的移植瘤体积和重量明显低于非特异siRNA转染组和空白对照组（P＜0.05）.STAT3 siRNA转染组移植瘤组织的STAT3 mRNA和蛋白表达水平均明显降低.结论 靶向STAT3的siRNA可以抑制乳腺癌MCF7细胞体内外的生长,STAT3有可能成为新的乳�Objective To observe the effect of signal transducers and activators of transcription 3 （STAT3） gene silence on the growth of breast cancer cell line MCF7 in vitro and in vivo and discuss the feasibility and effectiveness of STAT3 used as gene therapeutic target for breast cancer. Methods Human breast cancer cell line MCF7 cells were divided into 3 groups：mock control group, control group transfected with scrambled sequence siRNA, and experimental group transfectod with STAT3 siRNA. The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were examined by MTT method and flow cytometry. MCF7 cells treated with STAT3-siRNA were transplanted subcutaneously in nude mice and their tumorgenic ability was observed. The STAT3 mRNA and protein levels of the samples from nude mice of different groups were detected by semiquantity RT-PCR and Western blotting and compared. Results After treatment with STAT3-siRNA, STAT3 mRNA （0.327 ±0.020 vs. 1.035 ±0.050, 1.093 ±0.018） and ptotein （0. 153 ±0.006 vs. 1.320 ±0.033, 1. 374 ± 0. 022） levels in the MCF7 cells transfected with STAT3-siRNA were significantly lower than that in the two control groups （ P 〈 0.05）. MTT assay showed that after transfection of the STAT3-siRNA into MCF7 cells, cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was （44.00±5.10）%, significantly higher than that in the control group （16.1 ±1.05 ）% （P 〈 0. 05 ）. Flow cytometry results suggested that more apoptosis was observed in the STAT3-siRNA group. The apoptosis rate was （ 14.79±0.22）%, much higher than that in the control group [（7.06 ±0.71 ） %, （ 8.45 ± 0.43 ） %, P 〈 0.05]. The tumor growth in the experimental group was significantly slower than that in the two control groups. On the 22th day after transplantation, the tumor weight [（21.4 ±10.6） mg vs. （88.6±12.2） mg, （57.2 ±21.9） mg] an

关 键 词：乳腺肿瘤 RNA干扰 STAT3转录因子 MCF7细胞系 小鼠 裸 

分 类 号：R686[医药卫生—骨科学]