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目的构建人前脑腓肽原基因真核表达载体,将其转入NIH3T3细胞中表达,为晚期癌症的镇痛研究奠定基础。方法用PCR的方法获得人脑腓肽原基因,构建人前脑腓肽原基因真核表达载体pCDNA3.1(+)-PENK,用脂质体2000包裹转染NIH3T3细胞,G418筛选稳定表达后检测培养液中脑啡肽的含量。结果pCDNA3.1(+)-PENK真核表达质粒可以在NIH3T3细胞中高表达。结论pCDNA3.1(+)-PENK可作为肿瘤镇痛基因治疗的实验研究的有力工具。
OBJECTIVE: To construct eukaryotic expression vector of human preproendothelioma gene and transfer it to NIH3T3 cells for expression in the study of analgesia in advanced cancer. Methods The human procutofibrin gene was obtained by PCR. The eukaryotic expression vector pCDNA3.1 (+) - PENK was constructed and transfected into NIH3T3 cells with lipofectamine 2000. The expression of PCDNA3.1 (+) - PENK was detected by G418 selection. The content of enkephalin in culture medium. Results The pCDNA3.1 (+) - PENK eukaryotic expression plasmid was highly expressed in NIH3T3 cells. Conclusions pCDNA3.1 (+) - PENK can be used as a powerful tool in the experimental study of gene therapy of tumor analgesia.