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目的研究抗Ⅳ型胶原酶单链抗体(Fv)和力达霉素辅基蛋白(LDP)在大肠杆菌表达的融合蛋白(Fv-LDP)与力达霉素(LDM)的活性发色团AE形成的基因工程强化融合蛋白Fv-LDP-AE的生物学活性及体外抗肿瘤作用。方法采用酶联免疫吸附剂测定、免疫荧光细胞化学染色和明胶酶谱分析Fv-LDP-AE生物学活性;通过MTT法、Hoechst 33342和碘化丙啶共染、DNA ladder和侵袭实验探讨其抗肿瘤作用。结果 Fv-LDP-AE对人肝癌BEL-7402和人结肠癌HT-29细胞的免疫反应呈阳性,可与BEL-7402细胞特异结合,并呈剂量依赖性抑制人高转移性巨细胞肺癌PG细胞内Ⅳ型胶原酶分泌。Fv-LDP-AE对BEL-7402和PG细胞的增殖抑制作用比力达霉素强,可诱导BEL-7402细胞裂亡、凋亡,抑制BEL-7402细胞侵袭。结论 Fv-LDP-AE是一种以Ⅳ型胶原酶为靶点的、具有高效抗肿瘤活性的抗体药物。
Objective To study the activity of the active chromophore AE (Fv-LDP) and lidamycin (LDM) in the expression of Fv and LDP in Escherichia coli The resulting genetically engineered fusion protein Fv-LDP-AE biological activity and in vitro anti-tumor effect. Methods The biological activity of Fv-LDP-AE was analyzed by enzyme-linked immunosorbent assay, immunofluorescence cytochemistry and gelatin zymography. MTT assay, Hoechst 33342 and propidium iodide co-staining, DNA ladder and invasion assay were used to investigate the anti-Fv- Tumor action. Results Fv-LDP-AE showed positive immunoreactivity to human hepatocellular carcinoma BEL-7402 and human colon cancer HT-29 cells and specifically to BEL-7402 cells in a dose-dependent manner and inhibited the proliferation of human high metastatic giant cell lung cancer PG cells Type IV collagenase secretion. The inhibitory effect of Fv-LDP-AE on the proliferation of BEL-7402 and PG cells was stronger than that of doxycycline, which could induce the apoptosis and apoptosis of BEL-7402 cells and inhibit the invasion of BEL-7402 cells. Conclusion Fv-LDP-AE is an antibody with high anti-tumor activity targeting type IV collagenase.