pEGFP-N1-NPRL2真核表达载体的构建及其对人鼻咽癌细胞增殖的影响  

Construction of recombinant plasmid pEGFP-N1-NPRL2 and its effect on proliferation of nasopharyngeal carcinoma cells

作　　者：乔宇[1] 白荣平 盛玉彪 

机构地区：[1]冀中能源峰峰集团有限公司总医院耳鼻喉科,河北邯郸056200 [2]冀中能源峰峰集团有限公司总医院中医科,河北邯郸056200

出　　处：《现代肿瘤医学》2016年第15期2363-2365,共3页Journal of Modern Oncology

摘　　要：目的:构建pEGFP-N1-NPRL2真核表达载体并观察其对鼻咽癌细胞株CNE1体外增殖的影响。方法:提取CNE1细胞中总RNA,RT-PCR扩增NPRL2并克隆至pEGFP-N1载体,鉴定出阳性克隆送测序,以重组质粒转染CNE1细胞。通过Western blot检测转染细胞中NPRL2蛋白的表达,CCK-8法检测细胞增殖的变化。结果:成功构建了pEGFP-N1-NPRL2真核表达载体,Western blot法检测到NPRL2蛋白的表达,CCK-8法检测发现NPRL2能够明显抑制肿瘤细胞增殖(P<0.05)。结论:成功构建pEGFP-N1-NPRL2真核表达载体并转染至CNE1细胞,NPRL2可抑制肿瘤细胞的增殖。Objective： To construct the eukaryotic expression vector of pEGFP- N1- NPRL2 and its expression in human nasopharyngeal carcinoma cell line CNE1. Methods： Total mRNA was extracted from human nasopharyngeal carcinoma CNE1 cells,NPRL2 gene was obtained by RT- PCR and cloned into pEGFP- N1 vector,then the recombinant pEGFP- N1- NPRL2 plasmid was constructed and transfected into CNE1 cells by Lipofectamine 2000. The expression of NPRL2 in CNE1 cells was detected by qRT- PCR and Western blot. Results： Corrected construction of pEGFP- N1- NPRL2 was identified by double enzyme digestion and DNA sequencing. NPRL2 gene expressed by the transfected cells was testified by qRT- PCR and Western blot. Conclusion： The recombinant pEGFP- N1- NPRL2 plasmid has been constructed successfully and can inhibit CNE1 cells proliferation in vitro.

关 键 词：NPRL2 鼻咽癌 CNE1 增殖 

分 类 号：R739.6[医药卫生—肿瘤]