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利用逆转录病毒载体系统ＰＡ３１７／ＬＮＣ－ＩＬ－２所产生的含有ＩＬ－２基因及新霉素抗性基因（ｎｅｏｒ）的重组病毒，经共培养转染ＳＭＭＣ－７７２１肝癌细胞株。ＩＬ－２基因受巨细胞病毒早期启动子调控，转染后的细胞置于含新霉素类毒性药物Ｇ４１８（４００ｕｇ／ｍｌ）培养基内培养、筛选３０天，结果果转染率约为１～２％。对存活下来的转染有ＩＬ－２基因和新霉素抗性基因的细胞形态和生长情况进行观察，同时从水平及蛋白质水平对外源性基因的转录和翻译进行检测。结果表明：ＳＭＭＣ－７７２１肝癌细胞在ＩＬ－２基因转染前后形态及生长情况无明显变化；培养上清活性检测发现转染后为转染前的２０～３０倍；逆转录ＰＣＲ分析显示转染后的ＩＬ－２ｍＲＮＡ量明显高于转染前。研究表明我们已成功地将ＩＬ－２基因导入肝癌细胞并获得持续、稳定、高效表达。
The recombinant virus containing the IL-2 gene and the neomycin resistance gene produced by the retroviral vector system PA317/LNC-IL-2 was transfected into the SMMC-7721 hepatoma cell line by co-culture. The IL-2 gene is regulated by the early promoter of cytomegalovirus. The transfected cells are cultured in G418 (400 μg/ml), a neomycin-containing drug for 30 days. The transfection rate is approximately 1. ~ 2%. The cell morphology and growth status of the surviving transfected IL-2 gene and neomycin resistance gene were observed, and the transcription and translation of the exogenous gene were detected at the level and protein level. The results showed that the morphology and growth of SMMC-7721 liver cancer cells before and after IL-2 gene transfection had no significant changes; the activity of culture supernatant was found to be 20-30 times more than that before transfection, and the reverse transcription PCR analysis showed transfection. The amount of IL-2 mRNA was significantly higher after transfection. Studies have shown that we have successfully introduced the IL-2 gene into hepatoma cells and obtained sustained, stable and efficient expression.