抗EpCAM单克隆抗体治疗肝母细胞瘤的动物实验研究  

Targeting therapy of anti-EpCAM monoclonal antibody in hepatoblastoma animal model

作　　者：陈立才 李朋飞[2] 黄一敏[1] 刘江斌[1] 吕志宝[1] Chen Licai;Li Pengfei;Huang Yimin;Liu Jiangbin;Lv Zhibao(Department of General Surgery,Shanghai Children's Hospital,Shanghai Jiao Tong University,Shanghai 200062,China;Department of General Surgery,Children's Hospital of Zhengzhou University,Zhengzhou 471003,China)

机构地区：[1]上海交通大学附属儿童医院普外科,上海市200062 [2]郑州大学附属儿童医院普外科,河南省郑州市471003

出　　处：《临床小儿外科杂志》2019年第1期63-68,共6页Journal of Clinical Pediatric Surgery

基　　金：上海市卫生与计划生育委员会重点课题(编号:201440031)

摘　　要：目的观察抗上皮细胞粘附分子(epithelial cell adhesion molecule,EpCAM)单克隆抗体靶向治疗人肝母细胞瘤(hepatoblastoma,HB)荷瘤鼠模型的疗效。方法体外培养HB HuH-6细胞,采用免疫组化检测Hu H-6细胞中EpCAM的表达。将10只裸鼠皮下接种HuH-6细胞,待建立稳定的HuH-6细胞荷瘤鼠模型后,将荷瘤鼠等额随机分为两组:实验组经鼠尾静脉注射抗EpCAM单克隆抗体,剂量30 ug/kg,每天1次;对照组经鼠尾静脉注射等量生理盐水;两组均给药3周。停药1周后取荷瘤鼠肿瘤组织进行病理检查,测算肿瘤的重量及体积,并计算抑瘤率。结果 HuH-6细胞传代培养成功,免疫组化染色可见其细胞膜EpCAM呈阳性表达。裸鼠接种Hu H-6细胞6周后可形成肿瘤体,病理检查证实为HB,成功建立HB荷瘤鼠模型。实验组肿瘤重量及体积分别为(35. 6±2. 97) mg和(1. 12±0. 09) cm^3,对照组肿瘤重量及体积分别为(45. 8±4. 59) mg和(1. 37±0. 77) cm^3,差异均有统计学意义(P <0. 05)。实验组肿瘤组织EpCAM表达阳性率(28. 3±5. 4)%,对照组肿瘤组织EpCAM表达阳性率(39. 7±4. 4)%,差异有统计学意义(P <0. 05)。结论动物实验结果表明,抗Ep CAM单克隆抗体能够抑制HB荷瘤裸鼠肿瘤的生长,为HB靶向治疗提供了一定的实验基础和理论依据。Objective To explore the efficacy of anti-EpCAM(epithelial cell adhesion molecule)monoclonal antibody for hepatoblastoma(HB)in a nude mice model.Methods HB cell line of HuH-6 was cultured and the expression of EpCAM in HuH-6 examined.After HuH-6 cell was inoculated subcutaneously,all mice were randomly divided into treatment and control groups.In treatment group,anti-EpCAM monoclonal antibody was injected into mice by tail vein(30 ug/kg qd daily)for 3 weeks.In control group,the same volume of saline was applied.The expression of EpCAM from implanted tumor mass was detected by immunohistochemistry.Weight and volume of implanted tumor were measured and then tumor inhibition rate was calculated 1 week later.Results HuH-6 cell was successfully sub-cultured and the expression of EpCAM was positive on cell membrane.After 6-week cell implantation,mass was confirmed as HB by histopathological examination.Compared with control group,anti-EpCAM monoclonal antibody could significantly inhibit tumor growth in control group.Tumor weight and volume were smaller in treatment group than those in control group[tumor weight:(35.6±2.97)vs.(45.8±4.59)mg(P<0.05);tumor size:(1.12±0.09)vs.(1.37±0.77)cm^3(P<0.05)].The positive rate of EpCAM expression was(28.3±5.4)%in tumor tissue of experimental group and(39.7±4.4)%in tumor tissue of control group.The positive rate of EpCAM expression was lower in tumor tissue of experimental group than that of control group.And the difference was statistically significant(P<0.05).Conclusion Anti-EpCAM monoclonal antibody can inhibit the growth of HB tumor cells in a murine model.And it serves as a potential monoclonal targeting therapy for pediatric HB.

关 键 词：肝母细胞瘤 上皮细胞粘附分子 抗体 单克隆 靶向治疗 模型 动物 

分 类 号：R726[医药卫生—儿科] R735.7[医药卫生—临床医学]