鼻咽癌细胞株、瘤株中EBV-LMP1基因的检测及变异性分析

Analysis of the Variation of EBV-LMP 1 Gene in Transplantable Nude Mouse NPC Tumor Cell Strain(SUNT) and Its Corresponding Cell Line(SUNE)

采用ＰＣＲ法检测了不同代次的鼻咽癌细胞株、瘤株（ＳＵＮＥ／ＳＵＮＴ）中的ＥＢＶ－ＬＭＰ１基因片断，并分析其变异性。结果发现直到１３１代ＳＵＮＥ、３０代ＳＵＮＴ中仍然能检测到ＬＭＰ１基因片断，且在传代过程中ＬＭＰ１基因未发生变异。但与Ｂ９５－８中的ＥＢＶ－ＬＭＰ１基因相比，ＬＭＰ１基因发生了变异，表现为Ｘｈｏｌ、Ｍｓｐ酶切位点的消失和ＳＳＣＰ单链迁移率的不同。该结果提示变异的ＥＢＶ－ＬＭＰ１基因可能与ＳＵＮＥ／ＳＵＮＴ肿瘤的特性有一定的相关性。

The exon 1 fragment of EBV-LMP 1 gene was amplified from B 95-8 、Raji, SUNE/SUNT by PCR technique. The specificity of PCR products were confirmed by Dig-EBV-LMP 1 probe. The PCR products were cleaved by endonucleases Xhol and Msp. The results show that EBV-LMP 1-Exon 1 derived from B 95-8 has Xhol and Msp site, but EBV-LMP 1-Exon 1 derived from SUNE/SUNT lose Xhol and Msp site. PCR-SSCP technique was also applied to analyze the stability and variation of EBV-LMP 1-Exon 1 derived from different cell strains. The results showed that SSCP fingerprint of B 95-8 、Raji and SUNE/SUNT differed from each other, indicating the existence of genetic variation in EBV-LMP 1-Exon 1. Similar patterns of the SUNE/SUNT were obtained, indicaiing the stability of EBV-LMP 1 gene in SUNE/SUNT. The persistence and stability of varied EBV-LMP 1 gene in SUNE/SUNT suggest it may play an important role in maintaining the malignancy of SUNE/SUNT in vitro.