抗HER2单链抗体融合凋亡蛋白对SKOV3细胞的靶向杀伤

Expression of a secreted fusion protein consisting of anti-HER2 antibody and reversed caspase-3 and its suppression on growth of SKOV3 cells

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

AIM: To investigate the killing effect on SKOV3 cells by fusion protein HER2 specific antibody reversed caspase 3 in the secreted form. METHODS: The reversed human caspase 3 gene was subcloned into pCMV e23scFv PEII PEIII to construct recombinant eukaryotic expression vector pCMV e23scFv PEII revcasp 3 and transfect Jurkat cells. The secreted expression of the in culture supernant of transfected Jurkat cells was detected by ELISA. The suppression of the fusion protein on growth of SKOV3 cells was examined by co culture with supernant of transfected Jurkat cells. RESULTS: There was the secreted expression of the fusion protein in Jurkat cells. Expressed product had the activity of inducing SKOV3 cells to death. CONCLUSION: The fusion protein expressed in the secreted form can targeted towards SKOV3 cells and kill them.