Objective The pCMV-Tag2B-Flag tag was ligated to the human mitochondrial solute carrier protein 25A37 （SLC25A37）gene to construct a eukaryotic expression vector，and the effect of pcMV-Tag2B-FLAG tag on the function of human lung cancer H460 cells was detected. Methods The coding region fragment of SLC25A37 in human breast library was amplified by PCR， and the fragment and Flag vector were sliced with the restriction enzymes EcoR I and Xho I，and then the recombinant plasmid was formed after linking. Western Blot（WB）was used to identify the expression of target gene in cells. CCK-8 and clonal formation were used to detect the effect of SLC25A37 on the proliferation of lung cancer H460 cells. Wound healing assay was used to detect the effect of SLC25A37 on the migration ability of H460 cells. Results The constructed Flag-SLC25A37 eukaryotic expression plasmid was transfected into H460 cells and the fusion protein was successfully expressed. Overexpression of SLC25A37 could enhance the proliferation and migration of H460 cells. Conclusion The eukaryotic expression vector of human SLC25A37 gene carrying Flag tag could be expressed in human lung cancer H460 cells，and the gene could promote the proliferation and migration of H460 cells. SLC25A37 might be an oncogene and provide an important target for the treatment of lung cancer.