Construction of a recombinant lentiviral vector for VHL and VHL shRNA and its effect on proliferation and apoptosis of renal cell carcinoma cells
10.3969/j.issn.1673-4254.2015.03.07
- VernacularTitle:VHL慢病毒表达和干扰载体的构建及对肾癌细胞增殖和凋亡的影响
- Author:
Donglai SHEN
1
;
Xin MA
;
Yu ZHANG
;
Yu GAO
;
Xingtao LI
;
Liangyou GU
;
Huijie GONG
;
Shaoxi NIU
;
Xu ZHANG
Author Information
1. 中国人民解放军总医院泌尿外科肾脏疾病国家重点实验室
- Keywords:
lentiviral vector;
VHL gene;
proliferation;
apoptosis;
renal neoplasm
- From:
Journal of Southern Medical University
2015;(3):348-354
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines. Methods Lentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by LipofectamineTM 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry. Results The recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05). Conclusion VHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.
