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目的 研究三氧化二砷(As2O3 )对多发性骨髓瘤(MM)细胞基因表达的影响,探讨As2O3 治疗MM的分子机制。方法 将MMU266细胞分成对照组和实验组。取培养48h的2组细胞提取总RNA后分离mRNA,应用抑制性差减杂交法分离差异表达基因,并连接于pGEM TEasyVector,转化大肠杆菌JM109,构成差减cDNA文库,经蓝白斑筛选后挑选白色菌落,提取质粒,序列测定后进行同源性比较分析。结果 第1次差减分离出5个下调的基因:①氨基肽酶N;②人类肿瘤翻译控制蛋白1;③人类ATP合成酶A链;④信号识别颗粒A10; ⑤线粒体ATP合成酶/ATP酶亚单位6。第2次差减分离出4个表达上调的基因:①钙结合蛋白A10;②角质素6A;③相对分子质量为45×103 的含MIP重复成分的剪接因子;④多聚腺苷酸结合蛋白。结论 As2O3 可能通过改变上述基因的表达,发挥其抑制MM细胞增殖、促进凋亡和诱导分化的作用。
Objective To study the effect of arsenic trioxide (As2O3) on the gene expression of multiple myeloma (MM) cells and to explore the molecular mechanism of As2O3 in the treatment of MM. Methods MMU266 cells were divided into control group and experimental group. The total RNA was extracted from the two groups of cells cultured for 48h, and then the mRNA was isolated. The differentially expressed genes were isolated by suppression subtractive hybridization and ligated into pGEM TEasyVector. The recombinant plasmid was transformed into E. coli JM109 to construct a subtracted cDNA library. , The plasmid was extracted, the homology comparison analysis after sequencing. Results The first subtraction subtracted five down-regulated genes: ① aminopeptidase N; ② human tumor translation control protein 1; ③ human ATP synthase A chain; ④ signal recognition particles A10; ⑤ mitochondrial ATP synthase / ATPase Subunit 6. The second subtraction subtracted four genes that were up-regulated: ① calcium-binding protein A10; ② keratin 6A; ③ splicing factor with MIP repeats at a relative molecular mass of 45 × 103; ④ polyadenylate-binding protein . Conclusion As2O3 may play a role in inhibiting proliferation, promoting apoptosis and inducing differentiation of MM cells by changing the expression of above genes.