过表达PRDM14非小细胞肺癌肿瘤细胞株的构建  

Construction of Non-small Cell Lung Cancer CellsOverexpressing PRDM14

作　　者：卢亚楠 刘思珑 麻雨晴 刘梦琪 齐悦 LU Yanan;LIU Silong;MA Yuqing;LIU Mengqi;QI Yue(Beijing City University,Beijing 100094,China)

机构地区：[1]北京城市学院生物医药学部,北京100094

出　　处：《北京城市学院学报》2022年第5期31-35,共5页Journal of Beijing City University

基　　金：2021年北京城市学院“城市新星计划”训练项目—过表达PRDM14的非小细胞肺癌肿瘤细胞筛选(202111418016)。

摘　　要：本研究通过克隆人的PRDM14基因,构建过表达PRDM14的真核表达载体,转染NCI-H1975细胞后得到高表达PRDM14的非小细胞肺癌细胞株。以人胚胎干细胞cDNA为模板,PCR扩增PRDM14基因编码序列,将其插入pcDNA3.1真核表达载体,经双酶切和测序验证后,将重组质粒转染到NCI-H1975肺癌细胞中,采用Western blot检测目的蛋白表达情况。结果表明,pcDNA3.1-prdm14重组载体构建成功,且目的蛋白能够在NCI-H1975细胞中正常表达。构建的过表达PRDM14非小细胞肺癌细胞株可以为后续研究该基因对非小细胞肺癌的调控作用及非小细胞肺癌的靶向药物研究等提供实验材料。In this study,we cloned human PRDM14 gene,constructed an eukaryotic expression vector overexpressing PRDM14,and transfected NCI-H1975 cells to obtain a non-small cell lung cancer cell line with high expression of PRDM14.Using human embryonic stem cell cDNA as template,the coding sequence of PRDM14 gene was amplified by PCR and inserted into pcDNA3.1 eukaryotic expression vector,after double enzyme digestion and sequencing verification,the recombinant plasmid was transferred into NCI-H1975 lung cancer cells,and the expression of the target protein was detected by Western blot.The results showed that the recombinant vector pcDNA3.1-prdm14 was successfully constructed,and the target protein could be normally expressed in NCI-H1975 cells.The construction of non-small cell lung cancer cells overexpressing PRDM14 can provide experimental materials for the follow-up study,such as the regulatory effect of the gene on non-small cell lung cancer and the research of targeted drugs for non-small cell lung cancer.

关 键 词：PRDM14 真核表达载体 非小细胞肺癌 

分 类 号：Q784[生物学—分子生物学]