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

Abstract

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 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.

Keywords

• cd74-ros1
• fusion pcr
• nsclc
• migration
• proliferation