Construction of pcDNA3.1-CD74-ROS1 Recombinant Plasmid and Effect of CD74-ROS1 Expression on Proliferation and Migration of Lung Cancer Cells

WU Yazhou, LI Jia, ZHANG Lei
2018 Zhongliu Fangzhi Yanjiu  
Objective To construct the recombinant plasmid carrying CD74-ROS1 fusion gene, and explore its effect on migration ability of A549 cells. Methods CD74 and ROS1 genes were obtained through PCR. The CD74-ROS1 fusion gene was constructed by fusion PCR. We inserted the fusion gene into the eukaryotic expression vector pcDNA3.1, and transfected the pcDNA3.1-CD74-ROS1 recombinant plasmid into A549 cells. The expression of CD74-ROS1 was detected by real-time PCR or Western blot. The cell proliferation
more » ... cell proliferation was observed by MTT method. The cell morphological or cell migration ability were tested by crystal violet staining or wound healing assay. Results The CD74-ROS1 fusion gene was identified by double enzyme digestion and DNA sequencing. The expression of CD74-ROS1 protein was significantly increased after transfection. The overexpression of CD74-ROS1 enhanced the cell proliferation, changed the cell morphology and increased the ability of cell migration. Conclusion pcDNA3.1-CD74-ROS1 recombinant plasmid is successfully constructed and expressed stably in A549 cells. CD74-ROS1 overexpression could promote cell proliferation and the morphological changes and enhance the migration ability of lung cancer cells.
doi:10.3971/j.issn.1000-8578.2018.17.1486 doaj:15ec2d1ca5ab4a53bce4b5997c697f56 fatcat:2ks53rfxbzgqrcxlzua74j2o3m