Inhibition of hypoxia inducible factor 1αby siRNA-induced apoptosis in human retinoblastoma cells
10.3969/j.issn.1009-0754.2016.01.009
- VernacularTitle:siRNA抑制缺氧诱导因子-1ɑ表达对人视网膜母细胞瘤细胞增殖与凋亡的影响
- Author:
Lili LANG
1
;
Yu GAO
;
Rongrong GE
;
Jianmei CHEN
;
Ming JING
Author Information
1. 同济大学附属东方医院眼科
- Keywords:
Hypoxia inducible factor 1α;
siRNA;
Y-79 retinoblastoma cells
- From:
Journal of Navy Medicine
2016;37(1):22-26
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of hypoxia inducible factor-1ɑknocked down by small interfering RNA ( siR-NA) on cell proliferation , apoptosis and apoptotic pathways of human Y-79 RB cells.Methods Cell proliferation , HIF-1ɑmRNA and protein levels were measured by MTT , RT-PCR and Western Blot on human Y-79 RB cells under both normoxic and hypoxic conditions , respectively .siRNA knockdown against HIF-1ɑwas carried out to suppress the expression of HIF-1ɑ.Cell apotosis was determined by double staining cells with the annexin V-FITC and PI.Caspase activity was assessed by the fluorometric assay .Biostatistical analyses were conducted with SPSS 16.0 software package.Results A multifold increase in HIF-1ɑmRNA and protein levels were observed af-ter cells were exposed to the hypoxic environment at 0 h,4 h,8 h and 12 h.Both mRNA and protein levels of HIF-1ɑwere attenuated or abolished by transfection with siRNA H IF-ɑunder normoxic and hypoxic conditions .Futhermore,knockdown of HIF-1ɑcould enhance hy-poxia-induced slight increase of Bax/Bcl-2 ratio and activate caspase-9 and caspase-3.Conclusion This study was able to show that a knockdown of HIF-1ɑby siRNAH IF-1ɑresulted in a decrease in proliferation and induction of apoptosis in human Y-79 RB cells under both normoxic and hypoxic conditions .HIF-1ɑexpression may be a promising strategy for the treatment of human RB in the future .
