反义寡核甘酸类_反义；缺氧诱导因子-1；胶质瘤_

HIF-1α反义寡核苷酸体外对人脑胶质瘤细胞U251的作用

http://www.100md.com 《郧阳医学院学报》 2006年第3期

反义寡核甘酸类,反义；缺氧诱导因子-1；胶质瘤,,\],反义寡核甘酸类，反义；缺氧诱导因子-1；胶质瘤,1材料与方法,2结果,3讨论

     [摘要\] 目的:研究缺氧诱导因子1α(HIF1α)反义寡核苷酸(ASODN)转染对人胶质瘤细胞增殖及凋亡的影响,探讨以HIF1α为靶点进行反义寡核苷酸转染治疗胶质瘤的有效性。方法:设计合成针对HIF1α的寡核苷酸片段.实验分6组：空白对照组、脂质体组、正义组和反义组(1.0 μmol/l、2.5 μmol/l和5.0 μmol/l)。利用脂质体包裹瞬时转染人胶质瘤细胞系U251,MTT法检测细胞增殖;采用AO/EB染色及末端标记法(TUNEL)检测细胞凋亡的变化。结果:转染后各组细胞增殖抑制率分别为(1.30±0.05)%、(1.98±0.35)%、(2.47±0.48)%、(22.30±2.08)%、(33.60±4.19)%和(56.20±4.22)%。转染HIF1α反义寡核苷酸组较各对照组相比细胞增生下降, 凋亡增加(P<0.01)。结论:HIF1α反义寡核苷酸转染人胶质瘤U251细胞系可以抑制细胞增生并诱导胶质瘤细胞凋亡，HIF1α有望成为一个极具潜力的肿瘤治疗的靶点。

    [关键词\] 反义寡核甘酸类，反义；缺氧诱导因子-1；胶质瘤

    Effects of Antisense Oligodeoxynucleotides Targeting Hypoxia Inducible Factor 1α Gene on Human Glioma Cell Line U251 in Vitro

    YI Xian-fu,YANG Xian-yi,GUO Qing-hao,KANG Zhong-shan.

    (Emergency Department, Taihe Hospital,Yunyang Medical College,Shiyan,Hubei 442000,China)

    Abstract:Objective To investigate the effects of antisense oligodeoxynucleotides(ASODN) targeting HIF-1α on the apoptosis and proliferation of human glioma cell line U251 and to evaluate their efficacy as anticancer therapeutics. Methods ASODN targeting HIF-1α was designed and syntherized. Cultured U251 cells were devided into 6 groups:control group,lipsomes group,sense oligodeoxynucleotides(SODN) group,1.0 μmol/l ASODN group,2.5 μmol/l ASODN group and 5.0 μmol/l ASODN group. ODNs targeting HIF-1α transfected into U251 cells mediated by liposomal reagent. After transfected for 24 h , the cultured cells were harvested .Inhibitory rate(IR) by ASODN was determined by the colorimetri MTT cell viability;Apoptosis index (AI) and the apoptotic cell morphological changes were measured by acridine orange-ethidium bromide (AOEB) double fluorescent dye staining and TUNEL under fluorescence inverted microscopy. Results The IR in 6 group was (1.30±0.05)%,(1.98±0.35)%,(2.47±0.48)%,(22.30±2.08)%,(33.60±4.19)%,(56.20±4.22)%,respectively. The IR and AI rate in 3 ASODN groups was significantly higher than that in any other control groups(P<0.01). Conclusion ASODN targeting HIF1α may induce U251 apoptosis and inhibit proliferation. HIF1α could be a promising anticancer target . ......

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