摘要
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 investigate the effects of Chinese herbs of invigorating the spleen and removing blood stasis （JPHY）im- proving gene-therapy of human hepatocellular carcinoma（HCC） cell with adenovirus recombined by double suicide gene cytosine deami- nase（CD ）and herpessimplex virus thymidine kinase（TK ）, Methods： Adenovirus induced CD/TK double suicide genes were transfected into 293 cells by lipofectamine, The viral supernatant was collected. The human hepatocellular carcinoma BEL7402 cells were infected with the virus containing the double suicide genes, Cell survival rate was evaluated by MTT method, In the nude mouse HCC,CD/5-FC and HSV-tk/GCV systems were used together and JPHY herbs were administered through intragastric route, then the antitumor effects were evaluated according to tumor size, Results： The adenovirus containing CD/TK double suicide gene was produced in 293 cells. Double suicide genes were stably expressed in BEL7402 cells after being infected with the adenovirus. The cell killing effects of combined therapy with Chinese herbs and CD/TK genes group were superior to that of the CD/TK alone, In the mode of the nude mouse HCC, suicide gene therapy had strong inhibitory effect on tumor growth. The tumor size in the group treated with CD/TK＋5-FC/GCV combined with Chinese herbs was significantly less than that in the single gene therapy group or herbs group （P〈0.05）. Conclusion： The CD/TK suicide gene system has significant effect on killing 7402 cells. Chinese herbs of spleen invigoration and blood stasis removing can improve anti-tumor effects of CD/TK double suicide genes.
出处
《现代生物医学进展》
CAS
2008年第9期1605-1607,共3页
Progress in Modern Biomedicine
基金
中国博士后科学基金(No:2004035185)
广东省中医药局科研基金(No:1060034)
广州市卫生局科研基金(No:200221)
关键词
腺病毒
自杀基因
肝肿瘤
基阎治疗
中医药疗法
健脾化瘀药
Adenovirus
Suicide gene
Hepatocellular carcinoma
Gene therapy
Therapy of traditional Chinese medicine
Chinese herbs of spleen invigoration and blood stasis removing
