摘要
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 stromal cell-derived factor-1(SDF-1) gene expression in bone marrow stromal cells by a sequence-specific RNA degradation mechanism termed RNA interference. Methods Human SDF-1 specific short-interfering RNA expression plasmids were designed and synthesized, and were transfered into cultured human bone marrow stromal cells through cationic liposome-mediated gene transfer. Resistant clones were isolated by screening with G418 presence. SDF-1 mRNA and secreted SDF-1 protein of cells transfected with plasmid expressing short-interfering RNA (siRNA) were detected by reverse transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The cells transfected with plasmids expressing the same bases as siRNA but varied sequence at random and the untransfec- ted cells from the same samples were taken as control. Results Compared with untransfected and transfected with control plasmids cells, both of the SDF-1 mRNA and the SDF-1 protein of cells transfected with SDF-1-specific siRNA expression plasmids were decreased significantly. Conclusion The SDF-1 gene expression in human bone marrow stromal cells transfected with human SDF-1-specific siRNA expression plasmids is inhibited distinctly.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2005年第2期111-113,共3页
Immunological Journal
基金
国家自然科学基金资助项目 (3 0 170 3 96)
