[重组腺病毒介导的缺氧诱导因子1α突变在调节细胞凋亡中的机制]
[Mechanism of recombinant adenovirus-mediated mutations of hypoxia inducible factor 1alpha in modulation of cell apoptosis].
作者信息
Wei Li-li, Wu Ping-sheng, Wang Yue-gang, Hu Ying-fang, Xie Yi-jun
机构信息
Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2008 Mar;28(3):309-12.
OBJECTIVE
To investigate the mechanism by which recombinant adenovirus (Ad)-mediated mutations of hypoxia inducible factor 1alpha (Ad-HIF-1alpha-Ala564-Ala803) regulates cell apoptosis.
METHODS
LoVo cells were infected with recombinant Ad-HIF-1alpha-Ala564-Ala803 and control virus Ad-lacZ under normoxia condition. Real-time PCR was used to detect HIF-1alpha and p21WAF1/CIP1 mRNA expressions at different time points. Western blotting was employed to verify HIF-1alpha and p21WAF1/CIP1 protein expression. Hoechst 33342 flourescein staining was performed to observe the ratio of apoptotic LoVo cells.
RESULTS
The expression levels of HIF-1alpha mRNA and protein increased after infection with Ad-HIF-1alpha- Ala564-Ala803, accompanied by an increase in p21WAF1/CIP1 mRNA and protein expressions. The apoptotic ratio was significantly higher in LoVo cells infected with recombinant Ad-HIF-1alpha-Ala564-Ala803 (16.2%) than in the control cells (5.5%, P=0.00).
CONCLUSION
HIF-1alpha may induce cell cycle arrest by up-regulating p21WAF1/CIP1 expressions to promote LoVo cell apoptosis.
目的
研究重组腺病毒（Ad）介导的缺氧诱导因子1α（Ad-HIF-1α-Ala564-Ala803）突变调节细胞凋亡的机制。
方法
在常氧条件下，用重组Ad-HIF-1α-Ala564-Ala803和对照病毒Ad-lacZ感染LoVo细胞。采用实时荧光定量PCR检测不同时间点HIF-1α和p21WAF1/CIP1 mRNA表达。采用蛋白质印迹法验证HIF-1α和p21WAF1/CIP1蛋白表达。进行Hoechst 33342荧光染色观察凋亡LoVo细胞比例。
结果
感染Ad-HIF-1α-Ala564-Ala803后，HIF-1α mRNA和蛋白表达水平升高，同时p21WAF1/CIP1 mRNA和蛋白表达增加。重组Ad-HIF-1α-Ala564-Ala803感染的LoVo细胞凋亡率（16.2%）明显高于对照细胞（5.5%，P=0.0）。
结论
HIF-1α可能通过上调p21WAF1/CIP1表达诱导细胞周期阻滞，从而促进LoVo细胞凋亡。
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