Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the ...Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the use of immortalized cell lines in the laboratory to screen nucleic acid aptamers for recognition ability of tumor cells in the diseased body.To address this, primary cells that can be stably passaged were isolated and extracted from spontaneous tumors of genetically engineered pancreatic ductal adenocarcinoma model mice in this study.Next, an automated screening instrument for nucleic acid aptamers developed autonomously by our group was used to perform efficient aptamer screening using a limited number of cells, and the obtained nucleic acid aptamers were affinity verified at the cellular level.Finally, to answer the question of the cell growth environment difference on the recognition ability of nucleic acid aptamers, we verified its targeting ability to tumors in vivo on a nude mice xenograft tumor model, and further used a common antitumor drug doxorubicin combined with nucleic acid aptamers to verify the drug loading ability of this aptamer combined with the targeting therapeutic ability.展开更多
Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to dis...Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to discover some novel tumor markers,i.e.,membrane proteins that are specifically highly expressed on the surface of tumor cells.Tumor markers can be used in many fields such as early diagnosis and treatment,and a new type of tumor marker proved to be effective can significantly improve the therapeutic effect of such tumors.However,further characterization of newly acquired membrane proteins is essential for their clinical use as tumor markers.This review first briefly introduced the process of obtaining novel tumor markers from nucleic acid aptamers.Next,the commonly used protein characterization methods could be used as a technical means to identify membrane protein targets corresponding to tumor cell aptamers,to clarify the principles,advantages and disadvantages of various means,and to analyze the most suitable situations for various experimental methods.Finally,the outlook was made and the characterization methods that should be used in such experiments were summarized.展开更多
Since the concept of aptamer emerged,many scientists have launched a rich field of research around it.However,few nucleic acids aptamer which use cell as target can be put into practical applications.We believe that a...Since the concept of aptamer emerged,many scientists have launched a rich field of research around it.However,few nucleic acids aptamer which use cell as target can be put into practical applications.We believe that a great deal of this lies in the complexity and irreproducibility of aptamer screening experiments themselves.The complexity is due to the cumbersome processes and the technical requirements for laboratory personnel,whereas irreproducibility arises from the fact that the starting point of such screens is nucleic acid libraries with random fragments,and that different libraries directly determine the differences or even the success or failure of screening results.The complexity and irreproducibility mentioned above,in turn,lead to the inability of this experiment to unfold on a large scale,which naturally cannot lead to excellent results for practical applications.In response to this problem,our group has developed an instrument for automated screening of tumor cell nucleic acid aptamers and characterized the properties of nucleic acid aptamers obtained using this instrument in a comprehensive manner.展开更多
基金supported by the National Key Research and Development Program of China(Nos.2017YFA0205301 and 2018YFC1602905)National Natural Science Foundation of China(Nos.62071119,62075098,81902153,61527806)the Open Funding of State Key Laboratory of Oral Diseases(No.SKLOD2022OF05).
文摘Although it has been developed for many years, nucleic acid aptamer screening technology still fails to be widely used, a considerable part of it is due to the variability of tumor cell morphology, which leads to the use of immortalized cell lines in the laboratory to screen nucleic acid aptamers for recognition ability of tumor cells in the diseased body.To address this, primary cells that can be stably passaged were isolated and extracted from spontaneous tumors of genetically engineered pancreatic ductal adenocarcinoma model mice in this study.Next, an automated screening instrument for nucleic acid aptamers developed autonomously by our group was used to perform efficient aptamer screening using a limited number of cells, and the obtained nucleic acid aptamers were affinity verified at the cellular level.Finally, to answer the question of the cell growth environment difference on the recognition ability of nucleic acid aptamers, we verified its targeting ability to tumors in vivo on a nude mice xenograft tumor model, and further used a common antitumor drug doxorubicin combined with nucleic acid aptamers to verify the drug loading ability of this aptamer combined with the targeting therapeutic ability.
基金financially supported by the National Key Researchand Development Program of China(No.2018YFC1602905)National Natural Science Foundation of China(Nos.81902153,61871180,61527806)。
文摘Aptamer is an oligonucleotide chain with specific binding ability to protein and other targets,which is widely used in ma ny fields.Because of its ability to screen the premise of unknown targets,it can be used to discover some novel tumor markers,i.e.,membrane proteins that are specifically highly expressed on the surface of tumor cells.Tumor markers can be used in many fields such as early diagnosis and treatment,and a new type of tumor marker proved to be effective can significantly improve the therapeutic effect of such tumors.However,further characterization of newly acquired membrane proteins is essential for their clinical use as tumor markers.This review first briefly introduced the process of obtaining novel tumor markers from nucleic acid aptamers.Next,the commonly used protein characterization methods could be used as a technical means to identify membrane protein targets corresponding to tumor cell aptamers,to clarify the principles,advantages and disadvantages of various means,and to analyze the most suitable situations for various experimental methods.Finally,the outlook was made and the characterization methods that should be used in such experiments were summarized.
基金supported by the National Key Re-search and Development Program of China(No.2018YFC1602905)National Natural Science Foundation of China(Nos.6187118D,61527806)。
文摘Since the concept of aptamer emerged,many scientists have launched a rich field of research around it.However,few nucleic acids aptamer which use cell as target can be put into practical applications.We believe that a great deal of this lies in the complexity and irreproducibility of aptamer screening experiments themselves.The complexity is due to the cumbersome processes and the technical requirements for laboratory personnel,whereas irreproducibility arises from the fact that the starting point of such screens is nucleic acid libraries with random fragments,and that different libraries directly determine the differences or even the success or failure of screening results.The complexity and irreproducibility mentioned above,in turn,lead to the inability of this experiment to unfold on a large scale,which naturally cannot lead to excellent results for practical applications.In response to this problem,our group has developed an instrument for automated screening of tumor cell nucleic acid aptamers and characterized the properties of nucleic acid aptamers obtained using this instrument in a comprehensive manner.
