Construction and expression of a lentivirus expression vector carrying the VEGF165‑EGFP fusion gene in breast cancer MCF‑7 cells

Luo,Minna; Huang,Huan; Hou,Lei; Shao,Shan; Huang,Shangke; Zhao,Xinhan

doi:10.3892/ol.2017.5601

Construction and expression of a lentivirus expression vector carrying the VEGF165‑EGFP fusion gene in breast cancer MCF‑7 cells

Authors:
- Minna Luo
- Huan Huang
- Lei Hou
- Shan Shao
- Shangke Huang
- Xinhan Zhao
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Affiliations: Department of Oncology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Radiology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China, Department of Oncology, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China
Published online on: January 17, 2017 https://doi.org/10.3892/ol.2017.5601
Pages: 1745-1752

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Abstract

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 5.44x107 TU/ml in the LV‑VEGF165‑EGFP group and 5.00x108 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.

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