抗人大肠癌免疫毫微球的制备及其抗癌效果观察

PREPARATION AND ANTICANCER EFFECTS OF THE IMMUNONANOSPHERES AGAINST HUMAN COLORECTAL CANCER

目的 制备抗人大肠癌免疫毫微球 ,并观察其活性及疗效。方法 抗人大肠癌单克隆抗体SC3Ab通过异型双功能交联剂SPDP与载 5 Fu人血清白蛋白毫微球 [HSA(5 Fu) NS]偶联 ,制成抗人大肠癌免疫毫微球SC3Ab HSA(5 Fu) NS。使用凝集试验及免疫荧光检测其活性 ,光镜和电镜下观察其与大肠癌细胞SW 1116的特异性结合。MTT法检测该免疫毫微球的体外杀伤性。于人大肠癌裸鼠模型上分别使用SC3Ab HSA(5 Fu) NS、HSA(5 Fu) NS及 5 Fu ,检测三者的肿瘤抑制率。结果 SC3Ab HSA(5 Fu) NS具有单抗活性 ,能与大肠癌细胞特异结合 ;其体外杀伤SW 1116细胞IC50 值为 2 4 6 μg/ml ,与HSA (5 Fu) NS(345 3μg/ml)及 5 Fu(32 5 6 μg/ml)相比 ,明显降低 ;体内肿瘤抑制率比HSA(5 Fu) NS及 5 Fu明显增强 (P <0 0 0 1)。结论 SC3Ab HSA (5 Fu) NS具有免疫活性 ,对大肠癌细胞有主动靶向性 ,体内外均具有比HSA(5 Fu) NS及 5 Fu更强的抗癌效果。

Objective\ To prepare the immunonanospheres[SC3Ab HSA(5 Fu) NS] against human colorectal cancer and evaluate its immunoreactivity and effects on cancer. Methods\ SC3Ab HSA(5 Fu) NS was prepared by intermolecular cross linking the monoclonal antibody SC3Ab with human serum albumin nanospheres containing 5 Fu [HAS(5 Fu) NS] via new hetero bifunctional cross linker SPDP. Condensation test and immunoflurecence were used to evaluate the immunoreactivity, the specific binding of SC3Ab HSA(5 Fu) NS with colorectal cancer cell line SW1116 was observed by microscope and electron microscope. The specific cytotoxic effects on target cells were evaluated in vitro by MTT assay. SC3Ab HSA(5 Fu) NS, HSA(5 Fu) NS and 5 Fu were injected into nude mice bearing human colorectal carcinoma, to study the inhibitory activity of SC3Ab HSA(5 Fu) NS in vivo. Results\ The immunoreactivity of SC3Ab HSA(5 Fu) NS was well preserved. SC3Ab HSA(5 Fu) NS can bind the SW1116 cells specifically. The IC 50 value for SC3Ab HSA(5 Fu) NS on SW1116 cells was 24.6 μg/ml,which was lower than that of HSA(5 Fu) NS(345 3 μg/ml) and 5 Fu(325.6 μg/ml). The inhibitory rate of SC3Ab HSA(5 Fu) NS on the growth of colorectal cancer xenografts was significantly higher than that of HSA(5 Fu) NS or 5 Fu( P<0 001) . Conclusion\ SC3Ab HSA(5 Fu) NS has immunoreactivity and specific active targeting to the colorectal cancer cells. The anticancer ability of SC3Ab HSA(5 Fu) NS is significantly higher than that of HSA(5 Fu) NS and 5 Fu.