泰素免疫毫微球的制备及其抗人肝癌效果的动物实验研究

Preparation and anticancer effects of immunonanospheres containing paclitaxel against human liver cancer

目的制备抗人肝癌泰素免疫毫微球,观察其特性及抗人肝癌效果。方法抗人肝癌单克隆抗体HAb18通过异型双功能交联剂SPDP与泰素(Paclitaxel,PTX)人血清白蛋白毫微球[HSA(PTX)-NS]偶联,制成抗人肝癌泰素免疫毫微球HAb18-HSA(PTX)-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人肝癌细胞株SMMC-7721特异性结合。MTT法检测该免疫毫微球的体外杀伤性。于人肝癌裸鼠模型上分别使用HAb18-HSA(PTX)-NS、HSA(PTX)-NS及PTX,检测三者的肿瘤抑制率。结果 HAb18-HSA(PTX)-NS具有单抗活性,能与肝癌细胞特异结合;其体外杀伤SMMC-7721细胞IC50值为46.9μg/ml,与HSA(PTX)-NS(356.5μg/ml)及PTX(378.3μg/ml)相比,明显降低;体内肿瘤抑制率比HSA(PTX)-NS及PTX显著增强(P<0.001)。结论 HAb18-HSA(PTX)-NS具有免疫活性,对肝癌细胞有主动靶向性,体内外均具有比HSA(PTX)-NS及PTX更强的抗癌效果。

Objective To prepare immunonanospheres containing Paclitaxel[HAb18-HSA（PTX）-NS] against human liver cancer and evaluate its immunoreactivity and effects on cancer.Methods HAb18-HSA（PTX）-NS was prepared by intermolecular cross-linking the monoclonal antibody HAb18 with human serum albumin nanospheres containing PTX[HAS（PTX）-NS]via a new heterobifunctional cross-linker SPDP.Condensation test and immunofluorescence assay were used to evaluate the immunoreactivity,the specific binding of HAb18-HSA（PTX）-NS with liver cancer cell line SMMC-7721 was observed by microscope and electron microscope.The specific cytotoxic effects on target cells were evaluated in vitro by MTT assay.HAb18-HSA（PTX）-NS,HSA（PTX）-NS and PTX were separately injected into nude mice bearing human liver carcinoma,to study the inhibitory activity of HAb18-HSA（PTX）-NS in vivo.Results The immunoreactivity of HAb18-HSA（PTX）-NS was well preserved.Under electron microscope,HAb18-HSA（PTX）-NS could bind SMMC-7721 cells specifically.IC 50 value for HAb18-HSA（PTX）-NS on SMMC-7721 cells was 46.9μg/ml,which was lower than that of HSA（PTX）-NS（356.5μg/ml）and PTX（378.3μg/ml）.The inhibitory rate of HAb18-HSA（PTX）-NS on the growth of liver cancer xenografts was significantly higher than that of HSA（PTX）-NS or PTX（P0.001）.Conclusion HAb18-HSA（PTX）-NS has immunoreactivity and specific targeting to the liver cancer cells.The anticancer ability of HAb18-HSA（PTX）-NS is significantly higher than that of HSA（PTX）-NS and PTX.