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

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。

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

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 recombi-nant pEGFP -N1 -NPRL2 plasmid was constructed and transfected into CNE1 cells by Lipofectamine 2000.The ex-pression 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.