miRNA-21反义核酸对人淋巴瘤Raji细胞的生长抑制作用研究

为研究以miRNA-21为靶标的反义核酸（AMO-miR-21）对淋巴瘤Raji细胞的生长抑制作用,使用LipofectamineTM 2000将化学合成的反义核酸转染Raji细胞,采用四甲基偶氮唑蓝（MTT）法和台盼蓝拒染法检测细胞生长抑制率;实时定量PCR技术检测细胞miRNA-21水平改变;PI和Annexin V双染,流式细胞仪检测细胞凋亡.结果显示转染Raji细胞48～72h,反义核酸组细胞生长受到明显抑制,反义核酸抑制细胞活性的最佳浓度是0.4μmol/L;细胞内miRNA-21的表达水平明显下调;细胞凋亡明显增加;此外,反义组细胞中抑癌基因Pdcd4的mRNA和蛋白质呈高水平表达.提示以miRNA-21为靶标的反义核酸是人淋巴瘤Raji细胞生长的有效抑制剂和凋亡诱导剂.miRNA-21可能是淋巴瘤治疗的潜在靶标,其通过下调抑癌基因Pdcd4的表达水平发挥抗肿瘤作用.

Inhibitory Effects of AMO-miR-21 on Human Lymphoma Raji Cells

In order to study the inhibitory effects of anti-miRNA-21 oligonucleotides(AMO-miR-21) on human lymphoma Raji cells,human lymphoma cells(Raji cells)were transfected with chemically synthesized AMO-miR-21.The inhibition of cell growth was measured by MTT assay and trypan blue dye exclusion.miR-21 levels in cell were detected by quantitative real-time PCR.Apoptotic cells were evaluated flowcytometrically by double staining with Annexin V and Propidium Iodide.AMO-miR-21 obviously inhibited Raji cells growth and induced the cell apoptosis at the optimal inhibitory concentration of 0.4 μmol/L.AMO-miR-21 downregulated the expression level of miRNA-21.Meanwhile,mRNA and protein level of Pdcd4 were highly expressed in the Raji cells treated with AMO-miR-21.AMO-miR-21 maybe a strong Raji ceils growth inhibitor and apoptosis inducer.The mechanism of AMO-miR-21-mediated cell growth inhibition appears to be related to the direct modulation of downstream target Pdcd4,indicating that AMO-miR-21 might provide a potential targets for lymphoma therapy.