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将０．６ｋｂＬＭＰ１前段基因反向插入逆转录病毒载体（ｐＺＩＰ）中，构建成ｐＺＩＰ－反义ＬＭＰ１载体，再转入ＰＡ３１７包装细胞中，用Ｇ４１８筛选抗性克隆，获得产生１．７×１０５（ＣＦＵ／ｍｌ）反义ＬＭＰ１病毒的ＰＡ３１７细胞克隆。收集反义ＬＭＰ１病毒液，用于感染鼻咽癌细胞株（ＣＮＥ２）。杂交实验证实ｐＺＩＰ－反义ＬＭＰ１载体基因整合到ＣＮＥ２细胞的ＤＮＡ中，并且有载体基因ＲＮＡ的表达。观察反义ＬＭＰ１基因对ＣＮＥ２细胞生长的影响，发现反义ＬＭＰ１基因可降低细胞生长速度，减弱ＣＮＥ２细胞在裸鼠体内的致肿瘤性。原位杂交证实，肿瘤组织中持续存在着反义ＬＭＰ１基因。以上结果说明，ＥＢＶ反义ＬＭＰ１基因对ＣＮＥ２细胞的生长有明显的抑制作用。
Reverse the 0.6kb LMP1 gene into the retroviral vector (pZIP) to construct the pZIP-antisense LMP1 vector, transfer it to PA317 packaging cells, select the resistant clones with G418, and generate 1.7×105 ( CFU/ml) PA317 cell clones of antisense LMP1 virus. Antisense LMP1 virus fluid was collected for infection of nasopharyngeal carcinoma cell line (CNE2). Hybridization experiments confirmed that the pZIP-antisense LMP1 vector gene was integrated into the DNA of CNE2 cells, and there was expression of the carrier gene RNA. The effect of antisense LMP1 gene on the growth of CNE2 cells was observed. Antisense LMP1 gene was found to decrease the cell growth rate and reduce the tumorigenicity of CNE2 cells in nude mice. In situ hybridization confirmed the persistence of antisense LMP1 gene in tumor tissues. These results indicate that EBV antisense LMP1 gene significantly inhibited the growth of CNE2 cells.