CAJ | 학술논문

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72 h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.
배경:거서세포이동억제인자(MIF)시일충다공능세포인자,기재인체염증、면역반응화종류발생발전중기중요작용.목적:연구응용소간우RNA(siRNA)간우RNA(RNAi)기술조단MIF기인표체후대골육류세포주143B세포증식적억제효응.방법:전염MIF-siRNA위실험조,전염Fam-siRNA위대조조,채용면역형광법관측siRNA적전염솔,분별용q-PCR화Western blot검측143B세포중MIF-mRNA화MIF단백적표체수평,용MTT법검측143B세포증식능력.결과:실험조MIF-mRNA기인적상대표체량위대조조적26%,상비교유통계학차이(P<0.05);실험조MIF단백적상대표체량약위대조조적30%,상비교유통계학차이(P<0.05);143B세포전염72 h후,실험조능현저억제143B세포적활성.결론:MIF재골육류적발생발전과정중발휘중요작용,MIF-siRNA능조단MIF단백표체,종이억제143B세포증식,제시기가능성위골육류적신치료파점.
@@@@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-siRNA transfection group as the experimental group, and the Fam-siRNA transfection group as the com-parison 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 (P<0.05);and the relative gene expression quantity of MIF pro-tein in the experimental group was 30%higher than in the comparison group, there also existed statistical differences (P<0.05). 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 tar-get for osteosarcoma.