摘要
背景:巨噬细胞移动抑制因子(MIF)是一种多功能细胞因子,其在人体炎症、免疫反应和肿瘤发生发展中起重要作用。目的:研究应用小干扰RNA(siRNA)干扰RNA(RNAi)技术阻断MIF基因表达后对骨肉瘤细胞株143B细胞增殖的抑制效应。方法:转染MIF-siRNA为实验组,转染Fam-siRNA为对照组,采用免疫荧光法观测siRNA的转染率,分别用q-PCR和Western blot检测143B细胞中MIF-mRNA和MIF蛋白的表达水平,用MTT法检测143B细胞增殖能力。结果:实验组MIF-mRNA基因的相对表达量为对照组的26%,相比较有统计学差异(P<0.05);实验组MIF蛋白的相对表达量约为对照组的30%,相比较有统计学差异(P<0.05);143B细胞转染72h后,实验组能显著抑制143B细胞的活性。结论:MIF在骨肉瘤的发生发展过程中发挥重要作用,MIF-siRNA能阻断MIF蛋白表达,从而抑制143B细胞增殖,提示其可能成为骨肉瘤的新治疗靶点。
Background：Macrophage migration inhibitory factor（MIF） is a kind of pluripotent cytokine that plays an important role in human inflammation,immune responses,and tumor genesis and development.Objective：Adopting the small interfering RNA（siRNA） RNA interference（RNAi） technique to block the gene expression of MIF,and investigate its inhibitory effect on the proliferation of osteosarcoma cell line 143B cell.Methods：Take the MIF-siRNAtransfection group as the experimental group,and the Fam-siRNAtransfection group as the comparison group;observe the transfection efficiency by immunofluorescence technique;detect the expression level of MIF-mRNA and MIF protein in 143B cell by q-PCR and Western blot technique;and test the proliferation ability of 143B cell by MTT assay.Results：The relative gene expression quantity of MIF-mRNA in the experimental group was 26% higher than in the comparison group,there were statistical differences between the two groups（P0.05）;and the relative gene expression quantity of MIF protein in the experimental group was 30% higher than in the comparison group,there also existed statistical differences（P0.05）.72 hours later after the transfection,the viability of the 143B cells were significantly inhibited in the experimental group.Conclusions：MIF plays an important role in the genesis and development of osteosarcoma;the MIF-siRNA can block the expression of the MIF protein and consequently inhibit the proliferation of 143B cells,thus it can be a new therapeutic target for osteosarcoma.
出处
《中国骨与关节外科》
2013年第3期270-273,共4页
Chinese Journal of Bone and Joint Surgery
基金
江苏省自然科学基金(BK2010461)
