Construction of lentivirus expression vector containing transcription factor Twist gene and its effect on promoting epithelial to mesenchymal transition of breast cancer cells MCF-7

Abstract:

Objective：To construct lentivirus expression vector containing Twist gene and to explore its overexpression induced epithe－lial to mesenchymal transition（EMT） in MCF-7 of breast cancer cells. Methods：pcDNA3/myc-tagged-Twist plasmid was amplified by PCR and then cloned into pLVX-puro-vector. After the construction being transfected into HEK-293 cells，the supernatant was collected to infect the MCF-7 cells. The expression of Twist E-cadherin and Vimentin was analyzed by immunofluorescence and Western blot. The invasive ability was evaluated by Transwell invasion assay in MCF-7-Twist cells. Results：The pLVX-puro-myc-tagged-Twist was confirmed correctly by DNA sequencing. Compared with that of vector control cells，the expression of Twist，E-cad－herin and Vimentin was much higher and the invasive ability was much stronger in MCF-7-Twist cells（P<0.05）. Conclusions：The recombinant lentivirus vector expressing Twist is constructed and is efficaciously expressed in MCF-7 cells，which could significantly promote the invasion ability of MCF-7 cells through EMT.