E2F1-siRNA真核表达载体转染对人乳腺癌细胞系MCF-7生物学特性的影响  

Influence of E2F1-siRNA eukaryotic expression vector transfection on biologic characteristics of human breast carcinoma cell line MCF-7

作　　者：黄慧敏 李惠翔[2] 张红[2] 

机构地区：[1]河源市人民医院病理科,广东河源517000 [2]郑州大学第一附属医院病理科,郑州450052

出　　处：《中华实用诊断与治疗杂志》2014年第7期647-649,共3页Journal of Chinese Practical Diagnosis and Therapy

摘　　要：目的探讨转录因子E2F1表达对人乳腺癌细胞系MCF-7增殖及侵袭能力的影响。方法将构建并已鉴定的E2F1-siRNA真核表达载体(pSIREN-E2F1)和阴性对照载体(pSIRENC)利用脂质体介导转染技术转染人乳腺癌细胞系MCF-7,经嘌呤霉素筛选,得到稳定抑制E2F1表达的乳腺癌细胞(SIR组),并设阴性载体转染细胞(SIRC组)和未转染细胞(MCF-7组)为对照。3组细胞采用RT-PCR、Western blot法检测E2F1基因表达情况;采用Transwell侵袭实验、MTT实验检测细胞侵袭能力和增殖能力。结果建立了稳定抑制E2F1基因表达的乳腺癌细胞(SIR细胞);SIR组细胞E2F1mRNA、E2F1蛋白表达水平及细胞穿过膜细胞数(0.857±0.015、0.776±0.012、87.000±5.000)均明显低于SIRC组(1.563±0.031、1.512±0.011、143.000±11.000)和MCF-7组(1.544±0.018、1.494±0.013、152.000±9.000),差异均有统计学意义(P<0.05);培养第2天开始,SIR组细胞增殖能力明显低于SIRC组和MCF-7组(P<0.05)。结论采用siRNA技术抑制人乳腺癌细胞E2F1表达可明显抑制乳腺癌细胞的增殖能力,降低其侵袭能力,E2F1可作为乳腺癌发生过程中有价值的生物学指标。Objective To explore the influence of E2F1 expression on proliferation and invasiveness of human breast carcinoma cells. Methods The constructed and identified eukaryotic expression vector pSIREN-E2F1 and negative control vector pSIRENC were transfected into breast carcinoma cell line MCF-7 by using lipofectamine transfecting technique. Positive colonies （SIR group） were established after puromycin selection. The negative vector transfected cells （SIRC group） and untransfected cells （MCF-7 group） were as controls. The expressions of E2F1 mRNA and protein in three groups were examined by Western blot and RT-PCR technique. The invasive abilities and proliferation were examined by transwell invasion filter and MTT in three groups. Results Stable transfectants （SIR cells） were established. The expression levels of E2F1 mRNA and protein as well as the cell counts passing through theca cells were lower in SIR group （0.857±0. 015, 0. 776±0. 012, 87. 000±5. 000） than those in SIRC group （1. 563±0. 031, 1. 512±0. 011, 143.000±11.000） and MCF-7 group （1.544±0.018, 1. 494±0. 013, 152. 000±9.000） （P〈0.05）. The invasive abilities of breast carcinoma cells were significantly lower in SIR group than those in SIRC group and MCF-7 group from the second day of culture （P〈0.05）. Conclusions E2F1 siRNA can suppress the proliferation of breast carcinoma cells and decrease the invasive capability by suppressing the expression of E2F1. E2F1 may serve as a useful biologic marker for breast carcinoma progression.

关 键 词：乳腺癌 E2F1 RNA干扰 抑制 MCF-7细胞 

分 类 号：R737.9[医药卫生—肿瘤]