title: [In vitro selection of single strand deoxyribonucleic acid aptamers binding to cells from patients with acute myeloblastic leukemia].
abstract: OBJECTIVE
  To screen aptamers binding CD33+/CD34- cells from patients with acute myeloblastic leukemia M2 subtype (AML-M2).
  
  
  METHODS
  CD33+/CD34- cells from patients with AML-M2 were taken as targeted cells, CD33+/ CD34- cells from normal people were taken as anti-selecting cells, and aptamers in the single strand deoxyribonucleic acid (ssDNA) library were then selected repeatedly by cell-systematic evolution of ligands by exponential enrichment (C-SELEX) technology, and amplified by polymerase chain reaction (PCR) to generate sub-ssDNA library. During the experiment, PCR amplification with fluorescently labeled primer and flow cytometry were performed to analyze the aptamers'enrichment of sub-library, and the final round product of the sub-ssDNA library was cloned. After the sequencing, the primary and secondary structures of the aptamers were analyzed.
  
  
  RESULTS
  Electrophoresis indicated that the product of PCR amplification for each round subssDNA library was able to see a clear DNA band in the agarose gel. After 13 rounds of screening, the fluorescence intensity of the sub-ssDNA library binding the cells ranged from 2.14% to 51.12%, reaching a steady state at the 13th round. A total of 30 clones were selected and sequenced, 22 of which contained 1 of the 4 conserved sequences of AAGTA, TATCT, AGATG and AAATT in their primary structure, but the remained eight aptamers contained none of the conserved sequence. Secondary structure analysis indicated that four stem-loops and loop simulation convex structures existed in the aptamers.
  
  
  CONCLUSIONS
  C-SELEX technology can be used to screen the aptamers binding primary cells from patients with leukemia. The aptamers selected from the CD33+/CD34- cells from the patients of AML-M2 subtype might be used for the diagnosis and treatment for leukemia.
authors.affiliation: Department of Hematology, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Guangping
authors.initials: G
authors.lastname: Wang
authors.forename: Shuqin
authors.initials: S
authors.lastname: Zhang
authors.forename: Yajing
authors.initials: Y
authors.lastname: Xu
authors.forename: Minyuan
authors.initials: M
authors.lastname: Peng
authors.forename: Hui
authors.initials: H
authors.lastname: Yin
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.forename: Sanqin
authors.initials: S
authors.lastname: Tan
authors.forename: Fangping
authors.initials: F
authors.lastname: Chen
mesh_terms: D000293:Adolescent; D000328:Adult; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology; D015214:Antigens, Differentiation, Myelomonocytic / Q000235:genetics* / Q000276:immunology; D052157:Aptamers, Nucleotide / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000235:genetics*; D005260:Female; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D052156:SELEX Aptamer Technique; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000235:genetics / Q000276:immunology; D055815:Young Adult
chemical_list: D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D063268:Sialic Acid Binding Ig-like Lectin 3