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目的 通过单纯疱疹病毒胸苷激酶 (HSV tk)基因对荷大肠癌裸鼠肿瘤的直接转导及杀伤作用研究 ,建立有效的自杀基因抑制大肠肿瘤的治疗方法。方法 脂质体法将含HSV tk基因的逆转录病毒载体LNtk及空载体G1Na在Psi 2、PA3 17包装细胞中进行交叉包装、收获病毒上清。4 0只BALB/C裸鼠分成 4组并皮下注射LoVo细胞成瘤 ,将含tk和G1Na的病毒上清分别直接注射到 2组大肠肿瘤部位并予以腹腔注射 3 0 0mg/kg体重环氧鸟苷 (GCV) ;另 2组为未转基因对照组 ,其中予以腹腔注射GCV 1组 ,予以腹腔注射生理盐水 1组。治疗结束后将标本制成石蜡切片进行病理学分析。实验数据的统计学处理采用组间t检验。结果 各组动物成瘤性无明显差异。治疗结束时肿瘤标本的重量分别为 :转tk基因且予以GCV治疗组 (94±5 )mg,转空载体予GCV治疗组 (1687± 3 15 )mg,未转基因予以GCV治疗组动物 (1684± 3 0 1)mg,未转基因予以生理盐水腹腔注射组(164 9± 2 82 )mg,实验组肿瘤与各对照组相比 ,不仅观察到明显的肿瘤杀伤作用 (均为P <0 .0 1,n=10 ) ,而且存在明显的病理学改变。结论 HSV tk自杀基因有望成为大肠癌辅助治疗的有效手段之一。
Objective To study the direct transduction and cytotoxicity of herpes simplex virus thymidine kinase (HSV tk) gene on nude mice bearing colorectal cancer and to establish an effective suicide gene therapy for colorectal tumor. Methods The retroviral vector LNtk containing HSV tk gene and empty vector G1Na were cross-packaged in Psi 2 and PA3 17 packaging cells by liposome method, and the virus supernatant was harvested. 40 BALB / C nude mice were divided into 4 groups and subcutaneously injected with LoVo cells to make tumors. The virus supernatants containing tk and G1Na were directly injected into 2 groups of colorectal tumor sites and injected intraperitoneally with 300 mg / kg body weight of epoxy birds Glycosides (GCV). The other two groups were non-transgenic control group, which were injected intraperitoneally with GCV 1 group and intraperitoneally injected with normal saline group 1. After treatment, the specimens were made into paraffin sections for pathological analysis. Statistical analysis of experimental data using t-test between groups. Results There was no significant difference in tumorigenicity between groups. The weight of tumor samples at the end of treatment were as follows: tk gene transfected with GCV (94 ± 5) mg, GCV untreated (1687 ± 3 15) mg, GCV untreated (1684 ± (P <0.01). Compared with the control group, the tumors in the experimental group not only showed obvious tumor killing effect (all P <0.01) , n = 10), but also obvious pathological changes. Conclusion HSV tk suicide gene is expected to become an effective adjunct therapy for colorectal cancer.