HIF－1α反义寡核苷酸体外对人脑胶质瘤细胞U251作用.docVIP

HIF－1α反义寡核苷酸体外对人脑胶质瘤细胞U251作用 　 作者：易显富, 杨贤义, 郭清浩, 康中山 [摘 要] 目的:研究缺氧诱导因子1α（HIF1α）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HIF1α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HIF1α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HIF1α反义寡核苷酸组较各对照组相比细胞增生下降, 凋亡增加（Plt;0.01）。 结论:HIF1α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，HIF1α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 　　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 (AOEB) 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（Plt;0.01）. Conclusion ASODN targeting HIF1α may induce U251 apoptosis and inhibit proliferation. HIF1α could be a promising anticancer target . 　　Key words:Oligodeoxynucleotide,Antisense;Hypoxia inducible factor 1α;Glioma 　　恶性肿瘤在生长过程中,组织增生过快必然会造成局部组织缺氧和供能与耗能之间的不平衡[1]。缺氧诱导因子1（hypoxia inducible factor 1α, HIF1）是调节肿瘤细胞对缺氧等微环