【作者】 张焕铃； 王俊霞； 尤红煜； 郑龙； 连伟光； 刘树锋；

【Author】 ZHANG Huan-ling,WANG Jun-xia,YOU Hong-yu,ZHENG Long,LIAN Wei-guang,LIU Shu-feng (Molecular biology laboratory of Hebei Medical University,Hebei key lab of Laboratory Animal,Shijiazhuang 050017, Hebei,China)

【机构】 河北医科大学分子生物学研究室,河北省实验动物重点实验室；

【摘要】 目的观察AFP表达调控载体对AFP表达阳性肝癌模型靶向治疗作用。方法:以人肝癌HepG2(AFP阳性)、人肝癌SMMC772 1(AFP阴性)细胞于BALB/c-nu裸小鼠右腋皮下荷瘤,14d成瘤。将构建好的pAFP-EGFP和pAFP-P53-EGFP重组质粒于肿瘤内注射,通过免疫组化观察p53在HepG2肿瘤中的特异性表达及对肿瘤细胞的杀伤作用。结果:荷HepG2、SMMC7721细胞株裸鼠14日皮下肿瘤生长良好,且肿瘤直径均为1.0cm左右,经HE染色证实造模成功。同时,AFP免疫组化结果显示接种HepG2细胞的肿瘤组织AFP表达阳性,SMMC7721细胞的肿瘤组织AFP表达为阴性。经pAFP-EGFP和pAFP-P53-EGFP重组质粒治疗后,p53的免疫组化分析结果显示接种HepG2细胞的裸鼠pAFP-P53-EGFP治疗组p53表达量显著高于其他各组,且可见明显细胞凋亡现象。结论:含AFP基因调控序列的pAFP-P53-EGFP载体可专一性地作用于AFP阳性肝癌细胞,引起肝癌细胞周期阻滞和凋亡。更多还原

【Abstract】 Objective To observe the targeted therapy of expression vector containing AFP regulation sequence in AFP positive hepatoma model.Methods:HepG2(AFP + ),SMMC7721(AFP- )were inoculate defferent in BALB/c-nu nude mice right axillary subcutaneous.The tumor can be seen at 14d.The pAFP -EGFP and pAFP-P53-EGFP recombinant plasmid were injected intratumoral.The specific expression of p53 and cytotoxicity to HepG2 tumor was observed by immunohistochemistry.Results:The subcutaneous tumor of SMMC7721 cells and HepG2,grew well,and the tumor diameter was about 1.0cm,the HE staining confirmed the model.At the same time,AFP immunohistochemical results showed that AFP expressed in the tumor tissue of HepG2 cells inoculated,and there was no AFP expression in the tumor tissue SMMC7721 cells inoculated.After treatment,p53immunohistochemical analysis showed that inoculation of HepG2 cells in nude mice treated with pAFP-P53-EGFP p53 expression was significantly higher than those of the other groups,and the obvious apoptosis was seen.Conclusion:The pAFP-P53-EGFP vectors containing the AFP gene regulatory sequences can be specifically applied to AFP positive hepatocellular carcinoma cells,induced cell cycle arrest and apoptosis.更多还原