Mei Yu-Ping, Zhu Xiao-Feng, Zhou Jun-Min, Huang He, Deng Rong, Zeng Yi-Xin
Department of Experimental Research, State Key Laboratory of Oncology in Southern China, Cancer Center, Sun Yat-sen University, 651 DongFeng Road East, GuangZhou 510060, China.
Cancer Lett. 2006 Feb 8;232(2):189-98. doi: 10.1016/j.canlet.2005.02.010.
Epstein-Barr Virus (EBV) is closely associated with B cell malignancies. However, whether EBV appears to be absolutely required for cell proliferation and survival in lymphoma cells is still unknown. In this study, small interfering RNA (siRNA) targeting LMP1 was employed to investigate the effect of LMP1 on cell proliferation in EBV-positive lymphoblastoid B-cell line. A plasmid stable encoding 21-nt small RNA specifically and efficiently interfering LMP1 was constructed, resulting in a substantial loss of LMP1 mRNA and a significantly decreased LMP1 protein expression. Our data demonstrated that cell proliferation was completely inhibited and apoptosis was induced after knockdown of LMP1 gene in lymphoblastoid B-cell line. Also, we found that suppression of LMP1 caused downregulation of telomerase protein expression and decreased telomerase activity in lymphoma cells. In EBV-negative NPC cell line, transfection of plasmid expressing LMP1 greatly enhanced telomerase protein expression. Our results suggested that siRNA targeting LMP1 can induce apoptosis in EBV-positive lymphoma cells and is associated with inhibition of telomerase activity and expression. siRNA-directed LMP1 silencing may be of the therapeutic value for preventing and treating those EBV-associated tumors.
爱泼斯坦-巴尔病毒(EBV)与B细胞恶性肿瘤密切相关。然而,EBV对于淋巴瘤细胞的增殖和存活是否绝对必要仍不清楚。在本研究中,采用靶向LMP1的小干扰RNA(siRNA)来研究LMP1对EBV阳性淋巴母细胞B细胞系中细胞增殖的影响。构建了一个稳定编码特异性且高效干扰LMP1的21核苷酸小RNA的质粒,导致LMP1 mRNA大量丢失且LMP1蛋白表达显著降低。我们的数据表明,在淋巴母细胞B细胞系中敲低LMP1基因后,细胞增殖被完全抑制且诱导了细胞凋亡。此外,我们发现抑制LMP1会导致淋巴瘤细胞中端粒酶蛋白表达下调以及端粒酶活性降低。在EBV阴性的鼻咽癌细胞系中,转染表达LMP1的质粒极大地增强了端粒酶蛋白表达。我们的结果表明,靶向LMP1的siRNA可诱导EBV阳性淋巴瘤细胞凋亡,并与端粒酶活性和表达的抑制相关。siRNA介导的LMP1沉默可能对预防和治疗那些EBV相关肿瘤具有治疗价值。