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DNA aptamers from whole-serum SELEX as new diagnostic agents against gastric cancer

机译：全血清SELEX的DNA适体作为抗胃癌的新诊断剂

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Gastric cancer is still among the leading causes of cancer deaths worldwide. Despite the improvements in diagnostic methods, the status of early detection has not been achieved so far. Early diagnosis of gastric cancer may significantly improve the cure rate of patients. Therefore, a new diagnostic method is needed. In this study, subtractive SELEX was performed to screen gastric cancer serum-specific DNA aptamers by using gastric cancer serum and normal serum as the target and negative serum, respectively. Four highly specific aptamers generated for gastric cancer serum, Seq-3, Seq-6, Seq-19 and Seq-54, were developed using whole-serum subtractive SELEX technology with K _(d) of 128 ± 26.3 nM, 149 ± 23.6 nM, 232 ± 44.2 nM, 202 ± 25.6 nM, respectively. These generated aptamers showed higher specificities toward their target serum by differentiating normal serum but closely related other cancer serums. The selected four high affinity DNA aptamers were further applied to the development based on qPCR method for the early detection of gastric cancer. In addition, we performed MALDI-TOF MS followed by secondary peptide sequencing MS analysis for the identification of the aptamer binding proteins. Among these potential biomarkers, APOA1, APOA4, PARD3, Importin subunit alpha-1 showed a relatively high score probability. Therefore, these four ssDNA aptamers generated in our study could be a promising molecular probe for gastric cancer diagnosis.
机译：胃癌仍是全世界癌症死亡的主要原因之一。尽管诊断方法有所改进，但迄今为止尚未实现早期检测的状态。胃癌的早期诊断可以显着提高患者的治愈率。因此，需要一种新的诊断方法。在这项研究中，以胃癌血清和正常血清分别为靶标和阴性血清，进行了减法SELEX筛选胃癌血清特异性DNA适体。使用全血清消减SELEX技术开发了针对胃癌血清产生的四种高度特异性适体Seq-3，Seq-6，Seq-19和Seq-54，K _（d）为128±26.3 nM，149±23.6 nM，232±44.2 nM，202±25.6 nM。这些生成的适体通过区分正常血清但与其他癌症血清密切相关，对目标血清显示出更高的特异性。选用的四种高亲和力DNA适体进一步应用于基于qPCR的胃癌早期检测。此外，我们进行了MALDI-TOF MS，然后进行了二级肽测序MS分析，以鉴定适体结合蛋白。在这些潜在的生物标志物中，APOA1，APOA4，PARD3，Importin亚基α-1表现出较高的得分概率。因此，我们研究中产生的这四个ssDNA适体可能是胃癌诊断的有前途的分子探针。

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