title: [Effects of PTPN11 on the Biological Characteristics of AML Cells].
abstract: OBJECTIVE
  To detect the expression of PTPN11 gene in acute myeloid leukemia (AML) cell line and to explore the effects of PTPN11 over expressing on proliferation and apotosis of AML cell lines.
  
  
  METHODS
  The expression of PTPN11 in AML cell lines（HEL,U937, K562, KG-1, HL -60） was detected by RT-PCR, Q-PCR and Western blot. The PTPN11 gene was amplified by RT-PCR. PTPN11 DNA fragement and the lentiviral vector PCDH-CD513B were digested by BamHI and EcoRI, and then ligated by T4 DNA ligase. Recombinant lentivirus was generated by co-transfection of three-plasmids into 293FT cells using lipofectamine 2000. Then Q-PCR and Western blot were used to detect the expression of PTPN11 in the lentivirus infected HEL and U937 cells. The CCK-8 and Annexin V/7-AAD assays were performed to evaluate effects of PTPN11 on proliferation, apoptosis of HEL and U937 cells.
  
  
  RESULTS
  All 5 AML cell lines expressed the PTPN11 gene, restriction analysis and gene sequencing confirmed that recombinant lentiviral vector was successfully constructed. After transfection of cells with the lentivirus, the recombinant plasmid could stably up-regulate the expression of PTPN11. Analysis of the proliferation and apoptosis of transfected AML cells indicated that as compared with the control group, the OD values of over-expression group were significantly higher and the apoptotic rates were significantly lower (P<0.05).
  
  
  CONCLUSIONS
  PTPN11 is expressed in all the 5 AML cell lines. The lentiviral expression vector carrying human PTPN11 and the engineered HEL and U937 cell lines stably up-regulating PTPN11 gene expression are successfully obtained. Over-expression of PTPN11 promotes the proliferation of AML cell lines and inhibit then apoptosis.
authors.affiliation: Institute of Hematology, Xuzhou Medical College, Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
authors.forename: Jun-Shuai
authors.initials: JS
authors.lastname: Yue
authors.affiliation: Institute of Hematology, Xuzhou Medical College, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
authors.forename: Qing-Yun
authors.initials: QY
authors.lastname: Wu
authors.affiliation: Institute of Hematology, Xuzhou Medical College, Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
authors.forename: Ling-Yu
authors.initials: LY
authors.lastname: Zenge
authors.affiliation: Institute of Hematology, Xuzhou Medical College, Department of Hematology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China. E-mail:linhmd@163.com.
authors.forename: Kai-Lin
authors.initials: KL
authors.lastname: Xu
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D015470:Leukemia, Myeloid, Acute; D010957:Plasmids; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D014162:Transfection
chemical_list: C516724:PTPN11 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11