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In Vitro Cell Dev Biol Anim. 2010 Feb;46(2):107-13. doi: 10.1007/s11626-009-9257-7.
Nucleic acid aptamers are regarded as rivals for antibodies and as such are being investigated for their therapeutic potential. In the present work, it is shown that two different high-affinity DNA aptamers developed previously by Ferreira et al. against MUC1 antigen (designated MUC1-5TR-1 and MUC1-S1.3/S2.2) on MCF7 breast cancer cells can be linked to the first component of complement (C1q) via a biotin–streptavidin system and induce significant killing of MCF7 cells in vitro. Cell viability was assessed by Trypan blue uptake and absorbance at 590 nm of stained cells following buffer washes and lysis in 1% SDS. While the killing effect is demonstrable versus various controls, dependent on aptamer dose, and reproducible, it appears to kill maximally about half of treated MCF7 cells. Possible reasons for the marginal killing effect include antigenic shedding in vitro and membrane-bound complement regulatory proteins (mCRPs) on the cell surface such as CD46, CD55, and CD59 which act to inhibit complement-mediated lysis of cells. Future in vitro research could benefit from application of mCRP-specific aptamers in combination with anti-MUC1 aptamers to overcome surface protective mechanisms while attacking the plasma membrane of MCF7 cells or other MUC1-expressing cancer cells. However, in vivo such a combination could have deleterious effects on normal MUC1-expressing cells as well.
核酸适体被视为抗体的竞争对手,因此正在研究其治疗潜力。在本工作中,表明 Ferreira 等人先前针对 MCF7 乳腺癌细胞上的 MUC1 抗原(分别命名为 MUC1-5TR-1 和 MUC1-S1.3/S2.2)开发的两种不同高亲和力 DNA 适体可以通过生物素-链霉亲和素系统与补体的第一成分(C1q)连接,并在体外诱导 MCF7 细胞的显著杀伤。通过台盼蓝摄取和染色细胞在缓冲液洗涤和 1% SDS 裂解后的 590nm 处的吸光度来评估细胞活力。虽然杀伤作用可针对各种对照物,取决于适体剂量,并具有可重复性,但它似乎最多可杀死约一半的处理 MCF7 细胞。杀伤效果较差的可能原因包括体外抗原脱落和细胞表面上的膜结合补体调节蛋白(mCRP),例如 CD46、CD55 和 CD59,它们抑制补体介导的细胞裂解。未来的体外研究可以受益于 mCRP 特异性适体与抗 MUC1 适体的联合应用,以克服表面保护机制,同时攻击 MCF7 细胞或其他表达 MUC1 的癌细胞的质膜。然而,在体内,这种组合可能对正常表达 MUC1 的细胞产生有害影响。