增殖细胞核抗原反义核酸联合5-FU抑制肝癌细胞生长  

Inhibition of PCNA ASODN combining 5-FU on hepetocarcinoma cells

作　　者：吴志慧[1] 蒋建伟[1] 陈涛[1] 陈超[1] 丁文婷[1] 陈妙巧[1] 

机构地区：[1]暨南大学医学院生化教研室,广东广州510630

出　　处：《暨南大学学报（自然科学与医学版）》2008年第6期557-561,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广州市科技计划项目(2002J1-C0361);暨南大学本科科技创新项目(cx06075)

摘　　要：目的:探讨增殖细胞核抗原(PCNA)基因反义寡核苷酸(ASODN)联合5-FU对离体肝癌细胞及小鼠肝癌生长的抑制作用。方法:用5-FU、5-FU联合PCNA ASODN、5-FU联合聚乙烯亚胺(PEI)介导的PCNA ASODN(PEI-ASODN)分别转染人肝癌SMMC-7721细胞,改良MTT法(WST法)分析细胞生长增殖抑制效果;建立小鼠肝癌腋下移植瘤和腹水瘤模型,腋下移植瘤模型小鼠,隔日给药共5次,第13天杀小鼠,观察小鼠瘤重、瘤体积改变;腹水瘤模型小鼠,连续腹腔注射给药7次,记录小鼠平均存活天数,绘制小鼠存活曲线。结果:5-FU联合PEI-ASODN时,5-FU对肝癌细胞的IC50降低为0.165μg/mL,肝癌细胞对5-FU敏感性提高了49.7倍;肝癌腋下移植瘤小鼠各治疗组的瘤重、瘤体积均得到不同程度的抑制,其中以5-FU联合PCNA反义核酸组最高;对于腹水瘤小鼠,以5-FU和PCNA反义核酸联合治疗组平均生存时间最长。结论:5-FU联合PEI-ASODN能有效抑制肝癌细胞增殖,抑制小鼠肝癌的生长,延长小鼠存活时间。Aim： To evaluate the inhibitory effect of proliferating cell nuclear antigen antisense oligodeoxynueleotide combined with 5-FU on human hapetocarcinomar SMMC-7721 cells and carcinoma H22 cells transplated in mice. Methods： WST method was used to evaluate the proliferatory inhibition of SMMC-7721 cells and the IC50 using 5-FU only, 5-FU combined with PCNA ASODN, 5-FU combined with PCNA ASODN mediated by polyethyleneimine （PEI）. Transplanted hepatocarcinoma mouse model and ascites tumor mouse model with H22 cells were set up respectively. 5-FU was injected IP and ASODN was injected into the tumor tissues on the every other day after transplantation. On the 13th day, the mice were killed, the weights and volumes of subcutaneous tumors were measured and tumor inhibitory rates were calculated. Drugs were injected each day into the abdominal cavity during 1 week after H22 cells were inoculated, then the average survival time was calculated and the survival curve was plotted. Results： 5-FU inhibited the proliferation of SMMC-7721 cells when it was combined with PEI-ASODN, and the IC50 reduced to 0. 165 μg/mL. The sensitivity of SMMC-7721 ceils to 5-FU raised to 49.7 times.In the transplanted hepatocarcinoma mouse, the tumor inhibitory rate of 67.0% and the volume inhibitory rate of 72.4% were detected in the combined group, significantly different from the NS group（P 〈 0. 01 ）. In the mice ascites tumor model, the average survival time was 46.8 days and the life-prolong rate of 131% in the combined group when compared with the NS group, higher than other groups. In addition, a eooperativity effect was detected between 5-FU and PCNA ASODN. Conclusion： When 5-FU combined with PEI-ASODN, it can significantly inhibit the proliferation of hepatoeareinoma SMMC-7721 cells and the growth of mice hepatocareinoma and prolong the survival time of mice transplanted with hepatocarcinoma H22 cells.

关 键 词：肝癌 反义核酸 增殖细胞核抗原 5-氟尿嘧啶 

分 类 号：R736.7[医药卫生—肿瘤]