抗AGR2单克隆抗体18A4抑制细胞外AGR2诱导的肿瘤细胞活性

在两种自身不表达肿瘤相关蛋白AGR2的黑色素瘤细胞A375和B16-F10中加入AGR2以增强肿瘤细胞活性，观察以AGR2为靶点的单克隆抗体18A4对细胞外AGR2诱导的肿瘤细胞活性的抑制作用。用MTT法和克隆形成实验检测AGR2，18A4抗体单用和联合作用对细胞增殖能力的影响；流式细胞术检测AGR2，18A4抗体单用和联合作用对细胞周期的影响；划痕实验检测细胞迁移能力；鬼笔环肽染色检测细胞形态变化；Western blot和免疫荧光染色检测细胞内p53表达量。结果表明，单克隆抗体18A4能够抑制细胞外AGR2诱导的肿瘤细胞增殖速度加快，G₁/S细胞周期转化，迁移速度加快和细胞形态的变化。Western blot和免疫荧光染色检测发现在化疗药物阿霉素处理的细胞中，细胞外AGR2降低细胞内p53的表达量，而18A4能够抑制这一现象。实验结果表明18A4能够抑制细胞外AGR2诱导的多种肿瘤细胞活性，是一种潜在的抗肿瘤药物。

AGR2-blocking antibody 18A4 inhibits cellular tumorigenic properties induced by extracellular AGR2

This study was to evaluate the cellular inhibition effects induced by anti-AGR2 monoclonal antibody 18A4 in two AGR2-negative melanoma cells, A375 and B16-F10, treated with external oncoprotein AGR2. MTT assay and colony formation assay were performed to detect the cell proliferation rate. Flow cytometry analysis was performed to evaluate cell cycle transition. Cell migration rate was analyzed by wound healing assay. Cell morphological changes were detected by phalloidin staining. The expression of p53 was detected by Western blotting and immunofluorescence. Results showed that 18A4 inhibited the AGR2-dependent tumor properties including enhanced proliferation, accelerated cell cycle, increased cell migration and morphological changes of cells. In addition, by Western blot analysis and immunofluorescence, AGR2 blocking antibody 18A4 also restored p53 expression that was repressed by external AGR2 treated by chemotherapeutic drug doxorubicin. These findings suggest that 18A4 is able to inhibit the cellular tumorigenic properties induced by external AGR2 and is a potential drug against fumor.