XIAP反义寡核苷酸抑制胶质瘤细胞增殖和诱导凋亡的体外实验研究  

Antisense oligonucleotides targeting XIAP induces the apoptosis of malignant glioma cells

作　　者：张志清[1] 王晓芳[2] 

机构地区：[1]大连市中心医院神经内科,辽宁大连116033 [2]天津医科大学附属肿瘤医院内三科,天津300060

出　　处：《实用肿瘤杂志》2006年第4期297-301,共5页Journal of Practical Oncology

摘　　要：目的观察X连锁凋亡抑制蛋白(X IAP)反义寡核苷酸对人脑胶质瘤细胞系BT 325的增殖及凋亡的影响。方法将X IAP全硫代反义寡核苷酸,通过脂质体途径转染BT 325脑胶质瘤细胞。用CCK-8实验检测细胞杀伤率,RT-PCR和W estern b lot技术检测X IAP的mRNA和蛋白表达,JC-1检测细胞线粒体膜电位,流式细胞仪检测细胞凋亡率,比色法检测C aspase-3活性。结果X IAP反义寡核苷酸能够抑制BT 325脑胶质瘤细胞的增殖;X IAP反义寡核苷酸使X IAP的mRNA和蛋白表达均明显下调,mRNA较对照组减少了67.8%,蛋白较对照组减少了63.3%;反义寡核苷酸使线粒体膜电位下降,诱导BT 325细胞凋亡,反义寡核苷酸组的凋亡率为54.7%,错义链组为14.0%,两组比较,差异具有显著性(P<0.05);反义寡核苷酸使BT 325细胞C aspase-3活性明显增高,与对照组比较差异有显著性(P<0.05)。结论X IAP可能参与了脑胶质瘤细胞BT 325的增殖和凋亡,下调X IAP可抑制BT 325细胞增殖及促进其凋亡。Objective To investigate the effect of antisense oligonucleotides （AS ODNs） XIAP （X-linked inhibitor of apoptosis protein） on the proliferation and apoptosis of malignant glioma cells. Methods Fully phosphorothioated AS ODNs were delivered into malignant glioma BT325 cells with LipofectamineTM 2000 in the form of liposome-ODNs complexes. The colorimetric CCK-8 cell viability assay was used to detect the cytotoxicity of AS ODNs. Reverse transcription-PCR and Western blot were used to determine the expression of mRNA and protein of XIAP, respectively. Mitochondrial membrane potential was assessed using JC-1. Cellular Caspase-3 activities were observed by colorimetric method. Apoptosis was measured by flow cytometry. Results AS ODNs displayed a strong inhibitory effect on the proliferation of BT325 cells in a dose- and time-dependent manner. AS ODNs down-regulated the expression of XIAP mRNA by 67. 8% and XIAP protein levels by 63. 3%. The cell apoptosis rate in AS ODNs group was significantly higher than that in mismatch ODNs group （54.7% vs 14.0%, P〈0.05）. XIAP down-regulation resulted in Caspase-3 activation. Mitochondrial membrane potential of BT325 cells was decreased by AS ODNs. Conclusions XIAP might be involved in the proliferation and apoptosis of glioma cells. Down-regulation of XIAP might provide a new strategy for inhibiting the proliferation and inducing apoptosis of malignant glioma cells.

关 键 词：神经胶质瘤/病理学 脑肿瘤/病理学 寡核苷酸类 反义/药理学 细胞凋亡 基因表达 抑制蛋白类 肿瘤细胞 培养的 

分 类 号：R739.4[医药卫生—肿瘤]