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目的:观察健脾化瘀中药提高胞嘧啶脱氨酶/单纯疱疹病毒胸苷激酶基因治疗肝细胞癌的作用。方法:脂质体lipofectamine将含有双自杀基因的腺病毒载体pAd-CD/TK导人293细胞,收集病毒上清转染人肝癌细胞BEL7402,MTT法测定BEL7402细胞存活率。裸鼠人肝癌模型转染CD/TK双自杀基因后,给予5-FC500mg/kg,GCV 100mg/kg腹腔注射,同时予健脾化瘀中药960复方灌胃。观察肿瘤生长情况。结果:给予前体药物5-FC和GCV后,CD/TK转染细胞被杀死。并表现出较强的旁观者效应。转染细胞比例达到10%即表现出较强的杀伤作用(P<0.01)。健脾化瘀中药960复方具有提高旁观者效应作用,1.67ml/kg和2.5ml/kg960复方含药血清组细胞存活率显著低于对照组(P<0.01)。转染基因组应用5-FC和GCV治疗后,裸鼠肝癌的生长明显受到抑制(P<0.05),抑瘤率39.42%,单用中药组抑瘤率18.04%,中药与CD/TK+5-FC/GCV联合运用组,较单纯CD/5-FC/HSV-tk/GCV对裸鼠肿瘤模型的生长抑制作用更加明显(P<0.05),抑瘤率55.10%。结伦:腺病毒介导CD/TK自杀基因可有效地杀死人肝癌BEL7402细胞,健脾化瘀中药960复方具有显著提高CD/TK双自杀基因对人肝癌细胞的抑杀作用。
Objective: To observe the effect of invigorating the spleen and resolving phlegm medicine on cytosine deaminase/herpes simplex virus thymidine kinase gene therapy for hepatocellular carcinoma. METHODS: Liposomal lipofectamine was used to introduce human adenovirus vector pAd-CD/TK containing double suicide gene into human 293 cells. The virus supernatant was collected and transfected into human hepatoma cell line BEL7402. The survival rate of BEL7402 cells was measured by MTT assay. After the nude mice liver cancer model was transfected with CD/TK double suicide gene, 5-FC 500 mg/kg, GCV 100 mg/kg was injected intraperitoneally, and simultaneous invigorating spleen and removing blood stasis Chinese medicine 960 compound gavage. Observe the growth of the tumor. Results: After administration of the prodrug 5-FC and GCV, CD/TK transfected cells were killed. And showed a strong bystander effect. The proportion of transfected cells reached 10%, showing a strong killing effect (P <0.01). Invigorating the spleen and removing blood stasis Chinese medicine 960 compound has the effect of improving the bystander effect, and the survival rate of the 1.67 ml/kg and 2.5 ml/kg960 compound drug-containing serum group is significantly lower than that of the control group (P<0.01). After transfection with 5-FC and GCV, the growth of hepatocarcinoma in nude mice was significantly inhibited (P<0.05), the tumor inhibition rate was 39.42%, and the tumor inhibition rate was 18.04% in the Chinese herbal medicine group alone. Chinese medicine and CD/TK+5- Compared with CD/5-FC/HSV-tk/GCV, the inhibitory effect of FC/GCV on the tumor model of nude mice was more obvious (P<0.05), and the tumor inhibition rate was 55.10%. Conclusion: Adenovirus-mediated CD/TK suicide gene can effectively kill human hepatoma BEL7402 cells. Jianpi Huayu Chinese medicine 960 compound can significantly increase the killing effect of CD/TK double suicide gene on human hepatoma cells.