Objective: To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells.
Methods: A pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection.
Results: Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells.
Conclusion: Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.