Department of Cell Biology, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang North New Area, Shenyang 110122, Liaoning, China.
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, institute of chemistry, Chinese Academy of Sciences, 100190, Beijing, China.
Analyst. 2021 Dec 20;147(1):187-195. doi: 10.1039/d1an01579f.
Malignant melanoma is regarded as the most aggressive form of skin cancer, and is responsible for most death caused by skin cancer. mutations occur in approximately 40-50% of melanomas, with V600E being the most common mutation. Testing for mutations and targeted therapy have markedly improved long-term survival for patients with -mutated melanoma. Here, we report two aptamers, CH1 and CH5 generated by Cell-SELEX, against V600E-mutated human melanoma cells A375. The two aptamers exhibited high affinity to target cells with low dissociation constants () in the nanomolar range. Furthermore, the binding of two aptamers to target cells was independent of incubation temperature, and their molecular targets were demonstrated to be membrane proteins on the cell surface. We also demonstrated that aptamer CH1 was able to bind to metastatic colorectal cancer cells harboring V600E mutation, indicating a relationship between aptamer CH1 and V600E-related metastatic cancer. Owing to the structure stability and high selectivity to V600E-mutated targeting cells, aptamer CH1 holds great potential as a molecular probe for the detection of V600E-mutated metastatic melanoma.
恶性黑色素瘤被认为是最具侵袭性的皮肤癌，也是导致皮肤癌死亡的主要原因。大约 40-50%的黑色素瘤发生 突变，其中 V600E 是最常见的突变。检测 突变和靶向治疗显著改善了 -突变黑色素瘤患者的长期生存。在这里，我们报告了通过 Cell-SELEX 生成的针对 V600E 突变的人类黑色素瘤细胞 A375 的两个适体 CH1 和 CH5。这两种适体对靶细胞具有高亲和力，解离常数（）在纳摩尔范围内。此外，两种适体与靶细胞的结合不依赖于孵育温度，并且它们的分子靶标被证明是细胞表面的膜蛋白。我们还证明，适体 CH1 能够与携带 V600E 突变的转移性结直肠癌细胞结合，表明适体 CH1 与 V600E 相关的转移性癌症之间存在关系。由于结构稳定性和对 V600E 突变靶细胞的高选择性，适体 CH1 有望成为检测 V600E 突变转移性黑色素瘤的分子探针。
