AFP表达调控载体对肝癌模型的靶向治疗作用  

Targeted Therapy of Expression Vector Containing AFP Regulation Sequence in Hepatoma Model

作　　者：张焕铃[1] 侯洁[1] 王俊霞[1] 尤红煜[1] 郑龙[1] 连伟光[1] 刘树锋[1] 

机构地区：[1]河北医科大学实验动物学部,河北省实验动物重点实验室,石家庄050017

出　　处：《四川动物》2013年第4期605-608,共4页Sichuan Journal of Zoology

基　　金：河北省科技条件建设课题(No.12966121D);河北省自然科学基金(No.C2012206096);河北省卫生厅医学科学研究重点课题计划(No.20100039);河北省教育厅河北省高等学校科学研究指导计划(No.Z2010284)

摘　　要：目的观察甲胎蛋白(AFP)表达调控载体pAFP-P53-EGFP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SMMC7721(AFP阴性)细胞于BALB/c-nu裸小鼠右腋皮下荷瘤,14d成瘤,免疫组化检测AFP。将构建好的pAFP-EGFP和pAFP-P53-EGFP重组质粒于肿瘤内注射,观察肿瘤体积变化,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14d皮下肿瘤生长良好,且肿瘤体积均为500mm3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,可见明显细胞凋亡现象,且肿瘤体积较对照组减小。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。Objective To observe the targeted therapy of pAFP-P53-EGFP containing AFP regulation sequence in AFP positive hepatoma model. Methods HepG2 （AFP ＋ ）, SMMC7721 （AFP-） were inoculate in BALB/c-nu nude mice right axillary subcutaneous. AFP was detected by immunohistochemical. Tumor would be formed at 14 d. The pAFP-EGFP and pAFP-P53-EGFP recombinant plasmid were intratumorally injected and the tumor volume was monitored. The specific ex- pression of p53 and cytotoxicity to HepG2 tumor was observed by immunohistochem^stry. Results The subcutaneous tumor of SMMC7721 and HepG2 cells were grew well, and the tumor volume was about 500 mm3. The successfully constructed model was confirmed by HE staining. AFP immunohistochemical results showed that AFP expressed in the HepG2 cells in- oculated tumor tissue rather than SMMC7721. After treatment, p53 immunohistochemical analysis showed that expression of p53 in HepG2 cells inoculated nude mice treated with pAFP-P53-EGFP was significantly higher than other groups, and per- formed obvious apoptosis. The tumor volume was also smaller than the control group. Conclusion The pAFP-P53-EGFP vectors containing the AFP gene regulatory sequences, which is capable of inducing cell cycle arrest and apoptosis, can be specifically applied to AFP positive hepatocellular carcinoma cells.

关 键 词：AFP启动子 表达调控 P53 细胞周期阻滞 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]