基础医学与临床 ›› 2017, Vol. 37 ›› Issue (5): 630-635.

• 研究论文 • 上一篇    下一篇

长春新碱降低由转染A20 siRNA促进的弥漫大B细胞淋巴瘤细胞系增殖

王佳蕊1,王玲玲2,冯江龙1,杨文秀1   

  1. 1. 贵州医科大学病理学教研室
    2. 贵州医科大学
  • 收稿日期:2016-12-05 修回日期:2017-03-15 出版日期:2017-05-05 发布日期:2017-04-19
  • 通讯作者: 杨文秀 E-mail:ypq1964@163.com
  • 基金资助:
    国家自然科学基金资助项目

Vincristine reduces A20 siRNA-induced proliferation in diffuse large B cell

  • Received:2016-12-05 Revised:2017-03-15 Online:2017-05-05 Published:2017-04-19

摘要: 目的 研究A20小分子干扰RNA片段(siRNA)对弥漫大B细胞淋巴瘤细胞(OCI-LY1)增殖和MDR1表达的影响。方法 分对照组、转染组、长春新碱干预组(VCR)组和转染+VCR组。用脂质体转染法将A20 siRNA转入OCI-LY1细胞,MTT法检测细胞对VCR的敏感性;流式细胞计量术检测细胞凋亡;用real-time PCR检测A20及mdr1 mRNA;用Western blot检测细胞内A20、Pgp蛋白及核蛋白NF-κB的表达。结果 1. A20 siRNA转染后,细胞增殖能力增强(P<0.05),VCR刺激后转染细胞的增殖曲线下降缓慢。2.转染细胞凋亡率明显低于对照组,VCR刺激24h后OCI-LY1细胞凋亡率较加药前明显增加(P<0.05),VCR组凋亡率比转染+VCR组增高的幅度大(P<0.05)。3.转染后,A20mRNA及蛋白表达明显降低(P<0.001),NF-κB蛋白(P<0.001)、MDR1 mRNA(P<0.001)和Pgp (P<0.001)表达都明显增高;但MDR1mRNA 和Pgp在药物刺激后表达降低(P<0.05),且VCR组较转染+VCR组降低的幅度大(P<0.01)。结论 A20 siRNA能有效的增强OCI-LY1细胞内NF-κB的表达,使得MDR1基因及编码蛋白Pgp蛋白增强,从而抑制细胞凋亡、促进细胞增殖;VCR刺激后细胞内MDR1mRNA和Pgp的表达明显降低,A20 siRNA则减弱了VCR的这一作用。

关键词: 淋巴瘤, A20, NF-κB, MDR1, Pgp, RNA干扰, 细胞株

Abstract: Objective To investigate the impact of MDR1-targeting small interfering RNA (siRNA) on the proliferation of OCI-LY1 cells in diffuse large B-cell lymphoma. Methods A20 gene was silenced using RNA interference. An optimal concentration and treatment duration of vincristine were selected using MTT. Before and after siRNA transfection, proliferation of OCI-LY1 cells were assayed using MTT assay, and cellular apoptosis was detected using FCM before or after the treatment of the cells with VCR. Detection of A20, NF-κB (p65) and Pgp proteins were conducted using Western blotting whereas mRNA of the A20 and MDR1 genes were examined using real time PCR. Results 1. Proliferation of OCI-LY1 cells was enhanced (P<0.001) after the transfection with siRNA-2,(P<0.05). In addition, cell proliferation curve was declined after VCR stimulation, but the decrease was slower in siRNA-transfected cells than the untransfected counterparts. 2. Apoptostic rate was lower in siRNA-transfected cells than the untransfected counterparts, and the rate was higher in the cells after treatment with the drug for 24h (P< 0.05). Increased apoptosis was more obvious in common OCI-LY1 cells than in siRNA-transfected cells after the treatment with VCR(P<0.05). 3. The expression of MDR1 mRNA and Pgp (P <0.001) was significantly increased after transfection, but the expression of MDR1 mRNA and Pgp were significantly decreased (P<0.05). The expression in VCR group was significantly lower than that in siRNA-transfected cells+VCR group (P<0.01).C onclusion A20 siRNA could effectively enhance NF-kappa B expression in OCI-LY1 cells. NF-kappa B may up regulate the expression of its downstream genes such as MDR1 and cause apoptosis, in turn enhancing the inhibition of cell proliferation. VCR can reduce MDR1 mRNA and Pgp expression in OCI-LY1cells and the effect of VCR could be attenuated by A20 siRNA.