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目的 :探讨分泌表达的抗HER2单链抗体与重构型人caspase 3融合蛋白对HER2抗原阳性肿瘤细胞的靶向杀伤作用。方法 :将重构型人caspase 3基因亚克隆入pCMV e2 3scFv PEII PEIII的相应位点 ,构建重组真核表达载体 pCMV e2 3scFv PEII revcasp3,并转染人T淋巴瘤细胞系Jurkat,筛选并建系。用ELISA检测培养上清中融合蛋白的分泌表达。通过共培养实验观察含有该融合蛋白的培养上清对人卵巢癌细胞SKOV3生长的抑制作用。结果 :融合蛋白基因可在Jurkat细胞中 ,分泌表达并杀伤SKOV3细胞。结论 :分泌表达的抗erbB2单链抗体与重构型人caspase 3融合蛋白能够靶向诱导SKOV3细胞死亡
Objective: To investigate the targeted killing effect of secreted anti-HER2 single chain antibody and recombinant human caspase 3 fusion protein on HER2 antigen-positive tumor cells. Methods: The reconstructed human caspase 3 gene was subcloned into the corresponding sites of pCMV e2 3scFv PEII PEIII to construct recombinant eukaryotic expression vector pCMV e2 3scFv PEII revcasp3 and transfected into human T lymphoma cell line Jurkat, . Secretion of fusion proteins in culture supernatants was examined by ELISA. The inhibitory effect of the culture supernatant containing the fusion protein on the growth of human ovarian cancer cell SKOV3 was observed by co-culture experiments. Results: The fusion protein gene secreted and killed SKOV3 cells in Jurkat cells. CONCLUSIONS: The secreted secreted anti-erbB2 single chain antibody and the reconstructed human caspase 3 fusion protein are able to specifically target SKOV3 cell death