摘要
Liver cancer, as the second cause of cancer death all around the world, resulted in a series of chronic liver diseases. More than 80%of the patients cannot receive effective treatment because of their advanced disease or poor liver function. It is time to improve early clinical diagnosis and find optimal therapeutic treatments. As the tumor cells behave differently from the cell-surface molecules, it is necessary to find a highly specific probe. The aptamers, known as "chemical antibodies", can bind to their target molecules with high affinity and high specificity. The apatmers were obtained by Cell-SELEX, which was aimed at finding the aptamers against whole living cells. In the article, after 19 selections, the ssDNA pool was cloned and sequenced. After that, six aptamers were obtained, named apt_A to apt_F. By incubating the aptamers with different cells, except apt_E, the other aptamers showed high specificity. As for apt_E, which showed high affinity to several cancer cells, was a potential probe for the common protein presented by several different cancer cells. The equilibrium dissociation constants(Kd) were evaluated by measuring the flow cytometry signal that characterized the binding ability of aptamers to the target cells at a series of concentrations ranging from 46.3(4.5) nM to 199.4(44.2) nM, which exposed the high binding affinities of these aptamers. The research in the confocal fluorescence images further confirmed the specificity of these aptamers and the fact that the aptamers were combined with the targets on the cell-surface.
Liver cancer, as the second cause of cancer death all around the world, resulted in a series of chronic liver diseases. More than 80%of the patients cannot receive effective treatment because of their advanced disease or poor liver function. It is time to improve early clinical diagnosis and find optimal therapeutic treatments. As the tumor cells behave differently from the cell-surface molecules, it is necessary to find a highly specific probe. The aptamers, known as ＂chemical antibodies＂, can bind to their target molecules with high affinity and high specificity. The apatmers were obtained by Cell-SELEX, which was aimed at finding the aptamers against whole living cells. In the article, after 19 selections, the ssDNA pool was cloned and sequenced. After that, six aptamers were obtained, named apt_A to apt_F. By incubating the aptamers with different cells, except apt_E, the other aptamers showed high specificity. As for apt_E, which showed high affinity to several cancer cells, was a potential probe for the common protein presented by several different cancer cells. The equilibrium dissociation constants（Kd） were evaluated by measuring the flow cytometry signal that characterized the binding ability of aptamers to the target cells at a series of concentrations ranging from 46.3（4.5） nM to 199.4（44.2） nM, which exposed the high binding affinities of these aptamers. The research in the confocal fluorescence images further confirmed the specificity of these aptamers and the fact that the aptamers were combined with the targets on the cell-surface.
基金
supported by the National Science Foundation of China (61471168, 61571187, 61301043, 61527806)
the China Postdoctoral Science Foundation (2016T90403)
the Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province [(2013) 448]
