脂质体转染反义脱氧寡核苷酸对K562细胞α珠蛋白基因表达及细胞增殖的影响_医学论文.docVIP

脂质体转染反义脱氧寡核苷酸对K562细胞α珠蛋白基因表达及细胞增殖的影响_医学论文 脂质体转染反义脱氧寡核苷酸对K562细胞α珠蛋白基因表达及细胞增殖的影响_医学论文 【摘要】 本研究查明脂质体转染反义脱氧寡核苷酸（ASON）对K562细胞α珠蛋白基因表达的抑制作用及对细胞增殖的影响，探讨基因调控治疗β地中海贫血的新思路。 设计合成靶向α珠蛋白基因的ASON，并与正义寡核苷酸（SON）和空白对照进行比较。采用脂质体转染技术将ASON、SON与K562细胞共同培养，用荧光显微术、流式细胞术检测脂质体转染效率，用实时荧光定量RT法检测K562细胞中α珠蛋白基因表达情况，并通过Cell Counting Kit（CCK）法观察脂质体转染ASON技术对细胞增殖的影响。结果表明：脂质体转染效率在ASON作用24h达到最高，脂质体转染ASON组的α珠蛋白基因表达水平显著低于SON和空白对照组（plt0.01），并对细胞的增殖有明显的抑制作用，上述效应呈浓度依赖性。结论：脂质体转染ASON能特异性的抑制K562细胞中α珠蛋白基因表达，可能会成为β地中海贫血基因治疗的一个新靶点。 【关键词】 脂质体转染 Effect of Liposomal Transfection of Antisense Oligodeoxynucleotide on Alphaglobin Gene Expression and Proliferation of K562 Cells Abstract The objective of study was to investigate the effect of liposomal transfection of antisense oligodeoxynucleotide (ASON) on alphaglobin gene expression and proliferation of K562 cells, to explore the new way of gene therapy in βthalassemia. Targeted ASON of alphaglobin was designed and synthesized, and compared with positive control ［sense oligodeoxynucleotide (SON) group］ and blank control. By liposomal transfection, ASON, SON was coned by Cell Count Kit8 assay. The results indicated that the highest efficiency was at 24 hours after liposomal transfection, the gene expression level of alphaglobin in ASON group was significantly lower than that in SON group and blank control (plt 0.01). The proliferation of K562 cells was obviously inhibited, meanwhile the above effect showed the dosedependent manner. It is concluded that the liposomal transfection of ASON inhibits the alphaglobin gene expression of K562 cells, which may be the new target for gene therapy in βthalassemia. Key words alphaglobin gene liposome transfection antisense oligodeoxynucleotide K562 cell cell proliferation βthalassemia J Exp Hematol 2007 15(5):1065-1069 β地中海贫血是一种对人类健康危害严重的遗传性溶血性贫血病，其病理特征是β珠蛋白基因发生突变，引起β珠蛋白链合成减少，造成α与非α珠蛋白链的比例失衡，α珠蛋白链过剩，在红细胞内形成包涵体，导致骨髓无效红细胞生成和溶血。理论上基因治疗是最理想的方法，因而近20年来基因调控成为倍受人们关注的β地中海贫血治疗方法之一。目前针对β地中海贫血的基因调控治疗研究较多的停留在导入正常的β珠蛋白基因，或对剪接缺陷