^(131)I-CD133mAb联合顺铂在体内外有效抑制人肝癌细胞的生长

Inhibitory effect of ^(131)I-CD133mAb combined with cisplatin on liver cancer cells in vitro and in a tumor-bearing mouse model

目的观察131I-CD133mAb在体内外对高表达CD133抗原的Huh-7人肝癌细胞的靶向抑制作用。方法采用氯胺T法制备并鉴定131I-CD133mAb;流式检测Huh-7和HepG2细胞中CD133的表达,免疫组化检测瘤体内CD133的表达;实验设131ICD133mAb与顺铂(cisplatin,DDP)联用组、131I-CD133mAb组、DDP组及空白对照组。MTT法检测各组中不同药物处理对肝癌Huh-7细胞的生长抑制作用和计算每组药物的IC50值。成功构建人肝癌Huh-7细胞移植瘤模型,随机分为4组,1次/2 d给予尾静脉用药,连续2周,每周测量2次小鼠的体质量,测肿瘤的长径、短径。用药结束后处死小鼠,比较肿瘤的体积、质量,计算抑瘤率;HE染色法观察肿瘤组织病理学改变。结果131I-CD133mAb抗体标记率为90.25%,放射线化学纯度为97.78%;流式检测Huh-7细胞中CD133的表达为79.53%,明显高于HepG2细胞;免疫组化结果显Huh-7细胞CD133表达明显高于HepG2细胞;MTT法检测131I-CD133mAb+DDP组的细胞抑制率及体内抑瘤率明显高于131I-CD133mAb组、DDP组和空白对照组。结论131ICD133mAb+DDP在体内体外能有效地抑制高表达CD133抗原的肝癌细胞生长。

Objective To study the inhibitory effect of CD133 monoclonal antibody labeled with 131^I （131^I-CD133mAb） on Huh-7 human liver cancer cell line overexpressing CD133 antigen in vitro and in mouse models bearing the tumor cell xenograft. Methods 131^I-CD133mAb was prepared by chloramines-T method and evaluated for its stability. Flow cytometry and immunohistochemistry were used to detect the expression of CD133 in Huh-7 cells and in Huh-7 cell-derived tumors, respectively. Huh-7 cells treated with 131^I-CD133mAb plus cisplatin （DDP）, 131^I-CD133mAb, DDP, or no treatment （blank control） were examined for cell proliferation suppression by MTT assay with the IC50 calculated. BALB/c mice bearing subcutaneous Huh-7 cell xenograft in the right forelegs were treated with 131^I-CD133mAb, DDP, or both every two days for two weeks. The tumor size and volume were measured twice a week, and pathological examination of the tumor was carried out after the treatments. The tumor inhibition rate was calculated and tumor cell apoptosis observed with HE staining. Results The labeling ratio of 131^I-CD133mAb was 90.25% and the radiochemical purity was 97.78%. Huh-7 cells showed obviously higher CD133 expression than HepG2 cells. 131^I-CD133mAb combined with DDP group resulted in a significantly higher tumor inhibition rate than other treatments in the tumor-bearing mice. Conclusion 131^I-CD133mAb can inhibit the growth of liver cancer cells with a high CD133 expression both in vivo and in vitro.