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【目的】探讨前列腺特异膜抗原单克隆抗体7E11C5.3介导铼-188和平阳霉素(PYM)导向治疗前列腺癌细胞系LNCaP的体外作用。【方法】以葡聚糖为中介体制备7E11C5.3-PYM偶联物,间接ELISA法测定免疫活性,2,3,5-氯化三苯基四氮唑(TTC)法测定抑菌活性。采用2-巯基乙醇直接还原法制备188Re-7E11C5.3标记物,纸层析法测定标记率和放化纯度,直线回归外推法测定免疫活性分数。四唑盐(MTT)法测定体外细胞毒作用。【结果】7E11C5.3-PYM偶联物中7E11C5.3和PYM的克分子比为1∶54,偶联后单抗的免疫活性降低了10%～20%,偶联物中PYM的抑菌活性为游离PYM的25%。188Re-7E11C5.3的标记率为93.16%±2.18%,放化纯度为95.62%±0.48%,免疫活性分数为74.86%±1.86%。7E11C5.3-PYM和188Re-7E11C5.3对LNCaP细胞的抑制作用分别强于游离PYM和188ReO4-;两者单独用药时,其半数抑制浓度(IC50)分别为(10.17±2.06)μg/mL和(23.38±3.73)×104Bq/mL;两者联合用药时,其IC50值分别为(1.83±0.20)μg/mL和(6.82±0.73)×104Bq/mL。【结论】7E11C5.3-PYM和188Re-7E11C5.3联合用药对前列腺癌细胞的抑制作用显著强于单独用药。
【Aim】 To investigate the in vitro effects of 7E11C5.3 monoclonal antibody against prostate specific membrane antigen mediated by rhenium-188 and bleomycin (PYM) on prostate cancer cell line LNCaP. 【Method】 The 7E11C5.3-PYM conjugate was prepared with dextran as mediator. The immunological activity was determined by indirect ELISA. The antibacterial activity was determined by TTC method. 188Re-7E11C5.3 was prepared by 2-mercaptoethanol direct reduction method. The labeling rate and radiochemical purity were determined by paper chromatography. The linear regression extrapolation method was used to determine the immunological activity fraction. Tetrazolium salt (MTT) method for the determination of cytotoxicity in vitro. 【Result】 The molar ratio of 7E11C5.3 to PYM in 7E11C5.3-PYM conjugate was 1:54, and the immunogenicity of the conjugate decreased by 10% -20% after conjugation. The inhibition of PYM in the conjugate Activity is 25% of free PYM. The labeling rate of 188Re-7E11C5.3 was 93.16% ± 2.18%, the radiochemical purity was 95.62% ± 0.48%, and the immunological activity fraction was 74.86% ± 1.86%. The inhibitory effects of 7E11C5.3-PYM and 188Re-7E11C5.3 on LNCaP cells were stronger than that of free PYM and 188ReO4-, respectively. The IC50 of the two drugs were (10.17 ± 2.06) μg / mL and (23.38 ± 3.73) × 104Bq / mL, respectively. The IC50 values were (1.83 ± 0.20) μg / mL and (6.82 ± 0.73) × 104Bq / mL, respectively. 【Conclusion】 The combination of 7E11C5.3-PYM and 188Re-7E11C5.3 inhibited the proliferation of prostate cancer cells significantly better than that of single drug.