Gahn T A, Sugden B
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
J Virol. 1993 Nov;67(11):6379-86. doi: 10.1128/JVI.67.11.6379-6386.1993.
The Raji, EB1, and EB2 cell lines are derived from Epstein-Barr virus (EBV)-positive Burkitt's lymphomas. EBV gene products associated with viral latency, including latent membrane protein (LMP) and Epstein-Barr nuclear antigen 2 (EBNA-2), are expressed in these cell lines. We have found that transfection of all three of these cell lines by electroporation in both the presence and the absence of foreign DNA resulted in a marked decrease in expression of the endogenous EBV gene encoding LMP. An analysis of this response in Raji cells revealed that the level of RNA of this gene was decreased transiently and returned to normal levels by 7 days after transfection. The level of LMP protein was also reduced after transfection. No difference in survival was detected in electroporated versus unperturbed Raji cells. The level of mRNA encoding a modulator of the LMP gene, EBNA-2, was unchanged by electroporation. However, the level of another EBV transcript, BHLF-I, was reduced. The effect of electroporation could not be attributed to flux of Ca2+, Na+, K+, or Cl- ions across the plasma membrane. Expression of LMP in several lymphoblastoid cell lines was unaffected by electroporation.
Raji、EB1和EB2细胞系源自爱泼斯坦-巴尔病毒（EBV）阳性的伯基特淋巴瘤。与病毒潜伏相关的EBV基因产物，包括潜伏膜蛋白（LMP）和爱泼斯坦-巴尔核抗原2（EBNA-2），在这些细胞系中表达。我们发现，在有和没有外源DNA的情况下，通过电穿孔对所有这三种细胞系进行转染，导致编码LMP的内源性EBV基因的表达显著降低。对Raji细胞中这种反应的分析表明，该基因的RNA水平短暂下降，并在转染后7天恢复到正常水平。转染后LMP蛋白水平也降低。在电穿孔的Raji细胞与未受干扰的Raji细胞之间未检测到存活率的差异。编码LMP基因调节剂EBNA-2的mRNA水平不受电穿孔影响。然而，另一种EBV转录本BHLF-I的水平降低。电穿孔的影响不能归因于Ca2+、Na+、K+或Cl-离子穿过质膜的通量。几种淋巴母细胞系中LMP的表达不受电穿孔影响。
