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目的 通过RNA干扰来抑制骨髓基质细胞衍生因子 1(SDF 1)的表达。方法 利用脂质体介导人SDF 1特异性小片段双链RNA(siRNA)的表达质粒转染培养的人骨髓基质细胞,G4 18筛选阳性克隆。RT PCR和ELISA法检测培养细胞SDF 1mRNA和上清液SDF 1蛋白。以转染随机变更siRNA碱基序列的表达质粒和未转染的来源相同的骨髓基质细胞作为对照。结果 较未转染和转染随机变更siRNA碱基序列表达质粒的骨髓基质细胞,转染SDF 1特异性siRNA表达质粒的骨髓基质细胞SDF 1mRNA和SDF 1蛋白明显降低。结论 用SDF 1特异性siRNA的表达质粒转染骨髓基质细胞,抑制了SDF 1基因表达。
Objective To inhibit the expression of bone marrow stromal cell-derived factor 1 (SDF 1) by RNA interference. METHODS: Human bone marrow stromal cells were transfected with liposome-mediated expression plasmid of human SDF-1 specific double-stranded RNA (siRNA). Positive clones were screened by G418. The SDF 1 mRNA and SDF 1 protein of cultured cells were detected by RT PCR and ELISA. The expression plasmids with randomly modified siRNA base sequences were transfected into the untransfected bone marrow stromal cells with the same source as the control. Results Compared with untransfected and transfected BMSCs transfected with siRNA base sequence plasmid, SDF 1 mRNA and SDF 1 protein of bone marrow stromal cells transfected with SDF 1-specific siRNA expression plasmid were significantly decreased. Conclusion The expression of SDF-1 gene was inhibited by transfecting bone marrow stromal cells with SDF 1-specific siRNA expression plasmid.