摘要
背景与目的:微小RNA(microRNA,miRNA)是一类内源性的非编码小分子RNA,其主要通过与靶mRNA作用抑制转录后的翻译。miR-21是唯一一个在9种人类肿瘤细胞中高表达的小RNA分子。本研究旨在观察anti-miR-21反义寡核苷酸(ASO)对骨肉瘤MG-63细胞增殖的影响。方法:通过转染miR-21ASO和对照组ASO,采用实时荧光定量RT-PCR检测MG-63细胞中miR-21的表达变化,并运用多种细胞增殖检测方法观察miR-21ASO对MG-63细胞调控产生的系列效应。结果:与对照相比,miR-21ASO显著减少miR-21的表达(P=0.003);克隆形成实验结果显示miR-21ASO导致克隆形成率较对照组低(P=0.012);MTT实验结果显示转染miR-21ASO后,MG-63细胞存活数明显低于对照组(P=0.023);在体实验结果发现miR-21ASO抑制肿瘤细胞增殖,导致肿瘤质量较对照轻(P=0.013),形成的体积较对照小(P=0.028);流式细胞仪检测显示转染miR-21ASO后,MG-63细胞凋亡明显增加(P=0.022)。结论:miR-21ASO对骨肉瘤细胞miR-21水平降低有靶向特异性,转染miR-21ASO可有效抑制miR-21的促癌作用,利用反义核酸技术可为骨肉瘤的基因治疗开辟新的径路。
Background and purpose：MicroRNAs are small non-coding endogenous RNA molecules that can inhibit protein translation partly through binding with target mRNAs. MiR-21, as an oncomiR, is over expressed in 9 kinds of human tumor cells. The aim of this study was to investigate the regulative effects of miR-21 ASO on the proliferation of MG-63 cells. Methods：The miR-21 expression in MG-63 cells was measured using real-time PCR. Cell proliferation after transfection with miR-21 ASO was analyzed using MTT assay and the colony formation experiment in vitro and in vivo. The cell apoptosis was analyzed by Annexin V. Results：After transfection with miR- 21 ASO, miR-21 expression in MG-63 cells was decreased significantly. MTT assay showed that viability of MG-63 cells reduced greatly after transfection with miR-21 ASO. Colony formation significantly decreased after transfection with miR-21 ASO. In vivo tumors derived from MG-63 cells transfected with miR-21 ASO grew substantially slow in comparison to the negative control during the whole tumor growth period. Annexin V assay also indicated that transfection with miR-21 ASO promoted apoptosis greatly. Conclusion：miR-21 function can be effectively incited by miR-21 ASO. Down-regulation of miR-21 expression by miR-21 ASO can now be considered a new option in the treatment of miR-21 overexpressing cancers such as osteosarcoma, in future.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2010年第8期561-565,共5页
China Oncology
基金
国家自然科学基金(No:30672130)
