HIF-1α反义寡核苷酸体外对人脑胶质瘤细胞U251的作用

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

目的:研究缺氧诱导因子-1α(H IF-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H IF-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H IF-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(1.0μmol/l、2.5μmol/l和5.0μmol/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H IF-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:H IF-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,H IF-1α有望成为一个极具潜力的肿瘤治疗的靶点。

Objective To investigate the effects of antisense oligodeoxynucleotides （ASODN） targeting HIF - lα 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/1 ASODN group,2.5 μmol/1 ASODN group and 5.0 μmol/1 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 significandy 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 .