腺病毒5型E1A基因对乳腺癌细胞化疗药敏感性的影响  

Influence of E1A Gene of Adenovirus Type 5 on Sensitivity of Breast Cancer Cells to Chemotherapeutics

作　　者：杨新宇[1] 李群[2] 李向良[3] 房学东[1] 

机构地区：[1]吉林大学第二医院普通外科,长春130021 [2]吉林大学基础医学院病理生物学教育部重点实验室,长春130021 [3]吉林大学中日联谊医院泌尿外科,长春130021

出　　处：《中国生物制品学杂志》2007年第9期652-655,共4页Chinese Journal of Biologicals

摘　　要：目的探讨E1A基因对乳腺癌细胞化疗药敏感性的影响。方法双酶切质粒pUC119-E1A,获得E1A基因,连接入pEGFP-C1载体,构建真核表达质粒pEGFP-E1A。经脂质体介导转染人乳腺癌细胞系SK-BR-3细胞株,经筛选获得稳定表达E1A基因的转染细胞。RT-PCR检测E1A基因的mRNA转录水平。体外观察各组细胞软琼脂集落及对抗肿瘤药物顺铂的敏感性。结果RT-PCR结果显示,只有重组质粒pEGFP-E1A转染的细胞在约380 bp处有一特异性扩增条带,与未转染组和空载体转染组相比,转染E1A基因后,SK-BR-3肿瘤细胞在软琼脂中形成的集落明显变小,集落形成率明显降低,对顺铂的敏感性明显提高。结论已成功构建E1A基因表达质粒,并稳定表达了E1A基因,表达产物明显抑制乳腺癌细胞的生长,有效提高肿瘤细胞对化疗药物的敏感性,为其临床应用提供了理论和实验依据。Objective To explore the influence of E1A gene of adenovirus type 5 on the sensitivity of breast cancer cells to chemotherapeutics. Methods Digest previously constructed recombinant plasmid pUCll9-E1A with EcoR I and BamH I,and insert the obtained E1A gene into plasmid pEGFP-Cl. Transfect human breast cancer cell strain SK-BR-3 with the constructed eukaryotic expression vector pEGFP-E1A in mediation of liposome and screen the transfected cells for stable expression of E1A gene. Determine the transcription level of E1A gene by RT-PCR. Observe the growth of transfected cells on soft agar plate and their sensitivity to cisplatin. Resuits A gene fragment at a length of about 380 bp was amplified from the cells transfected with recombinant plasmid pEGFP-E1A. Compared with those of SK-BR-34 cells untransfected or transfected with empty vector, the colonies of E1A gene-transfected SK-BR-3 cells formed on soft agar plate were significantly small,the colony formation rate was significantly low,and the sensitivity to cisplatin increased significantly. Conclusion The recombinant plamsid for stable expression of E1A gene was successfully constructed. E1A gene inhibited the growth of breast cancer cells and increased their sensitivity to chemotherapeutics significantly,which provided theoretical and practical basis for its clinical application.

关 键 词：E1A基因 乳腺癌 基因转染 化疗药物 敏感性 

分 类 号：R737.9[医药卫生—肿瘤]