Construction of VEGF eukaryotic expression vector and its effects on proliferation and chemosensitivity in leukemia cells
- VernacularTitle:VEGF_(165)真核表达载体的构建及其对白血病细胞增殖和化疗药物敏感性的影响
- Author:
Chunxia JING
;
Huan ZHANG
;
Guang YANG
- Publication Type:Journal Article
- Keywords:
Vascular endothelial growth factors;
Eukaryotic expression vector;
K562 cells;
Homoharringto nine;
Fluorouracil
- From:
Chinese Journal of Pathophysiology
2010;26(3):533-538
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct eukaryotic expression vector of human vascular endothelial growth factor (VEGF) gene, and to explore the effects of VEGF gene on proliferation and chemosensitivity of leukemia cells. METHODS: The recombinant eukaryotic express plasmid pEGFP-C1-hVEGF_(165) was constructed with conventional gene engineering methods. The pEGFP-C1-hVEGF_165 was transfected into leukemia cell K562. The proliferation of the recombinant eukaryotic expression plasmid was analyzed by CCK8 method. The level of VEGF mRNA was detected by reverse transcription and quantitative polymerase chain reaction (RT-PCR). The expression level of VEGF protein was determined by ELISA in cell culture medium and cells. The cell cycle was detected by flow cytometry. RESULTS: VEGF_(165) eukaryotic expression vector was successfully constructed, confirmed by enzyme digestion and sequencing. Compared with the K562 cells transfected pEGFP-C1, the K562 cells transfected with pEGFP-C1-hVEGF_(165) grew faster and expressions of VEGF mRNA and protein increased significantly. In addition, K562 cells transfected with pEGFP-C1-hVEGF_(165) had relatively higher cell survival rate to chemotherapy drugs homoharringtonine(HHT) and fluorouracil(FU). CONCLUSION: VEGF_(165) eukaryotic expression vector increases the level of VEGF mRNA and protein expression, accordingly promotes the proliferation of leukemia cells and decreases the sensitivity of leukemia cells to HHT and FU.
