title: [Effects of stromal cells on sentitivity to imatinib in Sup-B15 Philadelphia chromosome positive acute lymphoblastic leukemia cells].
abstract: OBJECTIVE
  To investigate the sensitivity of imatinib (IM) on Sup-B15 Ph+ acute lmphoblastic leukemia (ALL) cells indused by stromal cells OP9, and to further explore its mechanism.
  
  
  METHODS
  The study is divided into two group, Sup-B15 cells group and co-cultured with OP9 cells group (Sup-B15/OP9 group). The inhibitory effects of IM on leukemia cells were measured by CCK-8 test, and the apoptosis by Annexin Ⅴ/7-AAD dyeing and the percentage of CD 34+CD38- leukemia cells were determined by flow cytometry. ALDH1, CD144, and β-catenin mRNA were detected by real-time RT-PCR, protein levels by Western blot. Inmunoprecipitation was used to detect the level of β-catenin connected to CD144.
  
  
  RESULTS
  IM presented inhibitory effects on Sup-B15 and Sup-B15/OP9 cells at multiple concentrations from 10 μmol/L to 45 μmol/L. The IC50 of IM on Sup-B15/OP and Sup-B15 cells were 35.8 μmol/L and 6.3 μmol/L, respectively (P<0.05). After 24 h of 30 μmol/L IM treatment, the percentages of apoptosis cells in Sup-B15/OP9 and Sup-B 15 cell were (14.24 ± 2.11)% and (3.45 ± 0.68)%, respectively (P<0.05). The percentage of CD34+CD38- cells in Sup-B15/OP9 group was significantly higher than that in Sup-B15 group [(3.42 ± 0.28)% vs (0.16 ± 0.15)%, P<0.05]. As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/OP9 group was remarkably upregulated (0.097 ± 0.012 vs 0.046 ± 0.010, P<0.05), and the CD133 protein level was also upregulated in Sup-B15/OP9 group. The transcription of CD144 in Sup-B15/OP9 group was remarkably upregulated compared with Sup-B15 group (0.103 ± 0.015 vs 0.010±0.003, P<0.05), as well as the CD144 protein. β-catenin mRNA transcription has no obvious changes between Sup-B15 group and Sup-B15/OP9 group (P>0.05), while the whole β-catenin protein and the cell nucleus β-catenin significantly increased, as well as the β-catenin protein combined with CD144.
  
  
  CONCLUSIONS
  Co-cultured with OP9 cells, Sup-B15 cells show less sensitivity to imatinib. The raising activity of CD144 and CD144/β-catenin signaling may work in this procession.
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Huanxin
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Zhiling
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Xuguang
authors.initials: X
authors.lastname: Song
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Depeng
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Qiu
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Pu
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Lingyu
authors.initials: L
authors.lastname: Zeng
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou　221000, China.
authors.forename: Kailin
authors.initials: K
authors.lastname: Xu
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D006801:Humans; D000068877:Imatinib Mesylate; D010677:Philadelphia Chromosome; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma; D015398:Signal Transduction; D017154:Stromal Cells; D051176:beta Catenin
chemical_list: C495270:CTNNB1 protein, human; D051176:beta Catenin; D000068877:Imatinib Mesylate