Open AccessConstruction and expression of a lentivirus expression vector carrying the VEGF165-EGFP fusion gene in breast cancer MCF-7 cells
Author(s) -
Min Luo,
Huan Huang,
Ling Hou,
Shan Shao,
Shangke Huang,
Xinhan Zhao
Publication year - 2017
Publication title -
oncology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.766
H-Index - 54
eISSN - 1792-1082
pISSN - 1792-1074
DOI - 10.3892/ol.2017.5601
Subject(s) - mcf 7 , lentivirus , oncogene , fusion gene , vector (molecular biology) , cancer research , molecular medicine , green fluorescent protein , viral vector , biology , expression vector , cell cycle , virology , microbiology and biotechnology , cancer , breast cancer , gene , human breast , human immunodeficiency virus (hiv) , genetics , viral disease , recombinant dna
Vascular endothelial growth factor (VEGF)165 is one of the most abundant and potent angiogenic factors in both physiological and pathological conditions. However, the function and mechanism of VEGF165 in tumors and their environment remain to be elucidated. In the present study, a lentivirus vector (LV) that contained the VEGF165-enhanced green fluorescent protein (EGFP) fusion gene was constructed and transfected into the human breast cancer cell line MCF-7. Following transfection, the expression of VEGF165 in MCF-7 cells was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. Further cellular localization of VEGF165 was observed through fluorescence microscopy. The titer of the recombinant lentivirus was 5.44×10 7 TU/ml in the LV-VEGF165-EGFP group and 5.00×10 8 TU/ml in the LV-EGFP negative control group. RT-qPCR and western blotting demonstrated that the expression of VEGF165 was significantly increased in the LV-VEGF165-EGFP group compared with the control group. The present study lays the foundation for in vitro and in vivo studies on tumor cell derived-VEGF165. Furthermore, the present fusion gene expression vector may provide a potential approach for gene therapy treatment of cancer and other diseases that require regulation of angiogenesis.