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目的 探讨重组免疫毒素scFv psm ETA对人前列腺癌细胞生长的抑制作用。 方法制备具有良好器官特异性的免疫毒素scFv psm ETA,从抗前列腺特异膜抗原(PSM)单抗杂交瘤细胞中克隆抗PSM单抗重、轻链可变区基因,与切去细胞结合区的ETA基因相连,构建成表达scFv psm ETA的质粒pSW200 psm,转化E.coli株CC118,表达有生物活性的融合蛋白scFv psm ETA,经M1FLAG柱纯化,检测其体外对前列腺癌细胞的细胞毒杀伤活性和在荷瘤裸鼠体内的抑瘤作用。 结果 制备的scFv psm ETA免疫毒素能特异性地与PSM高表达的LNCaP细胞结合,在浓度为100 ng/ml时对80%的LNCaP细胞有体外细胞毒杀伤作用;荷LNCaP前列腺癌裸鼠治疗组及对照组肿瘤大小分别为153mm3 及272mm3 (P<0. 01),显示scFv psm ETA对荷瘤裸鼠有抑制肿瘤生长作用。 结论 重组免疫毒素scFv psm ETA对PSM高表达的前列腺癌细胞LNCaP有体外细胞毒杀伤及体内抑瘤作用。
Objective To investigate the inhibitory effect of recombinant immunotoxin scFv psm ETA on the growth of human prostate cancer cells. Methods An immunotoxin scFv psm ETA with good organ specificity was prepared and cloned the anti-PSM monoclonal antibody heavy and light chain variable region genes from the anti-prostate specific membrane antigen (PSM) monoclonal antibody hybridoma cells, ETA gene to construct a recombinant plasmid pSW200 psm expressing scFv psm ETA. The recombinant plasmid pSW200 psm was transformed into E.coli CC118 and expressed as a biologically active fusion protein scFv psm ETA. The recombinant plasmid was purified on M1FLAG column to detect its cytotoxic activity against prostate cancer cells in vitro And in tumor-bearing nude mice in vivo anti-tumor effect. Results The prepared scFv psm ETA immunotoxin could specifically bind to LNCaP cells with high expression of PSM, and had cytotoxic effect on 80% of LNCaP cells at a concentration of 100 ng / ml. In addition, LNCaP prostate cancer nude mice treated with LNCaP The tumor size of control group was 153mm3 and 272mm3 respectively (P <0.01), which showed that scFv psm ETA could inhibit the growth of tumor-bearing nude mice. Conclusion The recombinant immunotoxin scFv psm ETA has cytotoxicity against LNCaP and its antitumor activity in vitro in PSM-overexpressing prostate cancer cells.