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目的研究缺氧诱导因子(HIF)1α反义寡核苷酸(ASODN)对人胶质瘤细胞化疗药物敏感性的影响。方法人工合成HIF1αASODN经阳离子脂质体包裹后瞬时转染人胶质瘤细胞株U251。采用RTPCR和免疫细胞化学检测转染后HIF1α基因表达,噻唑蓝(MTT)检测细胞增殖抑制率(PI),亚啶橙/溴化乙锭(AO/EB)染色及末端标记法(TUNEL)检测U251细胞转染后顺铂诱导的细胞凋亡指数(AI)。结果HIF1αASODN转染使U251细胞HIF1α基因表达明显下调,其AI和PI分别为(42.0±3.5)%和(72.5±4.8)%,,与各对照组比较均差异有统计学意义(P<0.01),而空白对照组,脂质体组和SODN组的AI和PI分别为(7.1±0.3)%,(8.2±0.2)%,(12.3±0.4)%和(30.7±2.9)%,(34.2±3.5)%,(38.6±3.1)%,AI和PI在各对照组之间的差异无统计学意义(P>0.05)。结论阳离子脂质体转染HIF1αASODN具有促进化疗药物诱导胶质瘤U251细胞凋亡及增强化疗药物敏感性作用。
Objective To investigate the effect of hypoxia inducible factor (HIF) 1α antisense oligonucleotide (ASODN) on chemosensitivity of human glioma cells. Methods Human HIF1αASODN was transiently transfected into human glioma cell line U251 by cationic liposomes. The expression of HIF-1α gene was detected by RTPCR and immunocytochemistry. The proliferation inhibition rate (PI), AO / EB staining and TUNEL assay were detected by MTT assay. Cisplatin-induced apoptosis index (AI) after transfection of U251 cells. Results The HIF1α ASODN transfection significantly down-regulated the expression of HIF1α gene in U251 cells, the AI and PI were (42.0 ± 3.5)% and (72.5 ± 4.8)%, respectively, which were significantly different from those of the control group (P <0.01) (7.1 ± 0.3)%, (8.2 ± 0.2)%, (12.3 ± 0.4)% and (30.7 ± 2.9)% in the blank control group, liposome group and SODN group respectively (34.2 ± 3.5%) and (38.6 ± 3.1)% respectively. There was no significant difference between AI and PI in each control group (P> 0.05). Conclusion Cationic liposome transfection with HIF1α ASODN can promote chemotherapeutic drugs to induce glioma U251 cell apoptosis and enhance chemosensitivity.