Suppression of Epstein-Barr nuclear antigen 1 (EBNA1) by RNA interference inhibits proliferation of EBV-positive Burkitt's lymphoma cells
Suppression of Epstein-Barr nuclear antigen 1 (EBNA1) by RNA interference inhibits proliferation of EBV-positive Burkitt's lymphoma cells
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DOI：
10.1007/s00432-005-0036-x
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Takano, Y
中科院分区：
文献类型：
--
作者：
Hong, M;Murai, Y;Takano, Y
目的：EB病毒(Epstein-Barr Virus，EBV)与包括Burkitt淋巴瘤在内的多种淋巴系和上皮性恶性肿瘤的发生发展有关。EBV潜伏蛋白EBV核抗原1(EBNA1)在几乎所有类型的EBV相关肿瘤中都可检测到，对EBV潜伏EB病毒的复制和维持至关重要。我们在这里研究了RNA干扰(RNAi)技术是否可以用来抑制EBV阳性的Burkitt淋巴瘤细胞中EBNA1的表达。方法：建立表达抗EBNA1小发夹RNA(ShRNAs)的Raji细胞系，用实时荧光定量RT-PCR方法检测EBNA1基因的表达水平。我们研究了EBNA1沉默对淋巴瘤细胞生长和细胞周期进程的影响。结果：与对照组相比，EBNA1 RNAi载体转染Raji细胞后，EBNA1基因表达缺失，Raji细胞增殖受到明显抑制。EBNA1的抑制还与EBV癌基因EBNA2的下调、增殖细胞核抗原标记指数的降低以及细胞周期分析中G0/G1比例的增加有关。结论：这些发现指出了载体介导的siRNA传递在控制EBV相关恶性疾病方面的潜在治疗应用。
Purpose: Epstein-Barr virus (EBV) is associated with the development of several lymphoid and epithelial malignancies, including Burkitt's lymphoma. The EBV latent protein, EBV Nuclear Antigen 1 (EBNA1), is detectable in almost all types of EBV-associated tumors and is essential for replication and maintenance of the latent episome of EBV. We here examined whether the RNA interference (RNAi) technique could be employed to suppress expression of EBNA1 in EBV-positive Burkitt's lymphoma cells. Methods: A Raji cell line expressing small hairpin RNAs (shRNAs) against EBNA1 was established and EBNA1 mRNA level was determined by real-time RT-PCR analysis. We investigated the effects of EBNA1 silence on lymphoma cell growth and cell cycle progression. Results: Transfection of an EBNA1 RNAi plasmid resulted in substantial loss of EBNA1 mRNA and significantly inhibited proliferation of Raji cells relative to the control plasmid case. Suppression of EBNA1 was also associated with downregulation of EBV oncogene EBNA2, a decreased PCNA labeling index and increased G0/G1 fraction in cell cycle analysis. Conclusions: These findings point to potential therapeutic applications for vector-mediated siRNA delivery to control EBV-associated malignant disorders.