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目的：了解ＨＳＶ－ｔｋ／ＧＣＶ系统协同放射治疗对小鼠黑色素瘤的联合杀伤作用。方法：通过逆转录病毒介导，将潮霉素磷酸转移酶和ＨＳＶ－ｔｋ的融合基因（ｈｙｔｋ）导入黑色素瘤细胞中并获得表达，通过体外和Ｃ５７ＢＬ／６小鼠动物实验，检测 ＧＣＶ对转基因细胞的体内外杀伤作用及联合应用放射治疗对黑色素瘤的治疗作用。结果：ＰＣＲ、ＲＮＡ Ｄｏｔ ｂｌｏｔｔｉｎｇ、免疫组化检测证实ＨｙＴＫ在转基因细胞的表达，体内外实验示ＧＣＶ对Ｂ１６／ＨｙＴＫ细胞有明显的杀伤作用，而对野生型Ｂ１６细胞无明显杀伤作用（Ｐ＜０．０５）；联合应用低剂量ＧＣＶ可使转入ｈｙｔｋ基因的黑色素瘤细胞对放疗的敏感性明显增高（ＳＥＲ＝１．６２），体内实验也证实协同疗法可延缓荷瘤小鼠的肿瘤生长。结论：ＨＳＶ－ｔｋ／ＧＣＶ系统协同放射治疗有望成为黑色素瘤基因治疗的一种有效方法。
OBJECTIVE: To understand the combined killing effect of HSV-tk / GCV system on melanoma in mice. Methods: The hytk gene of hygromycin phosphotransferase and HSV-tk was transfected into melanoma cells by retrovirus-mediated transfection and the expression was obtained. The effects of GCV on transgene In vitro and in vivo cytotoxicity of cells and combined treatment of melanoma with radiotherapy. Results: The expression of HyTK in transgenic cells was confirmed by PCR, RNA Dot blotting and immunohistochemistry. In vitro and in vivo experiments showed that GCV had a significant killing effect on B16 / HyTK cells and no significant cytotoxicity on wild-type B16 cells (P <0. .05). Combination of low-dose GCV can significantly increase the sensitivity of melanoma cells transfected with hytk gene to radiotherapy (SER = 1.62). In vivo experiments also confirmed that synergistic therapy can delay the tumor growth in tumor-bearing mice. Conclusions: Synergistic radiotherapy with HSV-tk / GCV system is expected to be an effective method for gene therapy of melanoma.