Abstract
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 Kd 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.
Highlights
Gastric cancer is the second most common malignancy in the world, threatening seriously the human health.[1]
The effective gastric cancer tumor biomarker get poor in clinical diagnosis and prognosis
A washing buffer solution containing 1 L of 1Â PBS, 1 mL of 1.5 mmol MgCl2 and the Tris phosphate-buffered saline (TPBS) solution was a mixture of 1 L of 1Â PBS with containing 1 mL of 1% (v/v) Tween-20, were stored at 4 C
Summary
Gastric cancer is the second most common malignancy in the world, threatening seriously the human health.[1]. Carcinoembryonic antigen (CEA) and CA125 are the commonly used tumor biomarkers in clinical practice for gastric cancer screening; early diagnosis, targeted treatment and prognosis are the main strategies to promote the ve-year survival rate.[2,3,4,5] the clinical diagnostic accuracy of CEA is unsatisfactory due to low sensitivity and speci city Another major challenge for gastric cancer is the lack of therapeutic options. Addition, we performed MALDI-TOF MS followed by secondary peptide sequencing MS analysis for the identi cation of the aptamer binds proteins Among these potential biomarkers, APOA1, APOA4, PARD3, Importin subunit alpha-1 showed a relatively high score probability. These four DNA aptamers generated here could be a promising molecular probe for gastric cancer diagnosis
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