title: [Effect of VE-cadherin on sensitivity to Imatinib in Sup-B15 Philadelphia chromosome positive acute lymphoblastic leukemia cells].
abstract: OBJECTIVE
  To investigate the sensitivity of imatinib mesylate (IM) on Sup-B15 Ph⁺ acute lymphoblastic leukemia (ALL) cells knockdown of VE-cadherin (CD144), and to further explore its mechanism.
  
  
  METHODS
  CD144 in Sup-B15 leukemia cells was stably knock downed via lentivirus-mediated RNA interference (named as Sup-B15/shVEC). The inhibitory effects of IM on Sup-B15/shVEC and Sup-B15 leukemia cells were measured by CCK-8 test, and the apoptosis of those cells was determined by AnnexinV/7-AAD dyeing using flow cytometry, the percentage of CD34⁺CD38⁻ leukemia cells also by flow cytometry. ALDH1 mRNA levels were detected by real-time RT-PCR, and protein levels of CD144, CD133, Bcr-abl and β-catenin by Western blot.
  
  
  RESULTS
  IM treatment presented inhibitory effects on Sup-B15/shVEC and Sup-B15 leukemia cells at multiple concentrations of IM. The IC50 of IM on Sup-B15/shVEC and Sup-B15 leukemia cells were 25.1μmol/L and 18.7μmol/L, respectively (P<0.05). After 48h of 20 μmol/L IM treatment, the percentages of apoptosis cell in Sup-B15/shVEC cells and Sup-B15 cell were (13.52±2.06)% and (3.03±0.72) %, respectively (P<0.05). The percentage of CD34⁺CD38⁻ cells in Sup-B15 cells was significantly higher than in Sup-B15/shVEC cells [(2.39±0.28)% vs (0.96±0.07)%, P<0.05). As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/shVEC was remarkably downregulated, and the CD133 protein level was also downregulated in Sup-B15/shVEC cells. Both cytoplasmic and nucleic β-catenin protein levels (but not for Bcr-abl levels) decreased in Sup-B15/shVEC cells as compare to Sup-B15 cells.
  
  
  CONCLUSIONS
  Knockdown of CD144 sensitized Sup-B15 Ph+ ALL cells to IM. The possible mechanisms underlying this phenomenon might be via inhibiting β-catenin nucleic translocation and facilitating β-catenin degradation.
authors.affiliation: Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Laboratory of Transplantation Immunology, Xuzhou 221000, China.
authors.forename: Huan-xin
authors.initials: HX
authors.lastname: Zhang
authors.forename: Zhi-ling
authors.initials: ZL
authors.lastname: Yan
authors.forename: Xu-guang
authors.initials: XG
authors.lastname: Song
authors.forename: Chao
authors.initials: C
authors.lastname: Lü
authors.forename: Jiang
authors.initials: J
authors.lastname: Cao
authors.forename: Zhen-yu
authors.initials: ZY
authors.lastname: Li
authors.forename: Ling-yu
authors.initials: LY
authors.lastname: Zeng
authors.forename: Chong
authors.initials: C
authors.lastname: Chen
authors.forename: Kai-lin
authors.initials: KL
authors.lastname: Xu
mesh_terms: D015703:Antigens, CD / Q000235:genetics; D001549:Benzamides / Q000494:pharmacology*; D015820:Cadherins / Q000235:genetics; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D000068877:Imatinib Mesylate; D010879:Piperazines / Q000494:pharmacology*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology*; D034622:RNA Interference; D051176:beta Catenin / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D001549:Benzamides; D015820:Cadherins; D000068877:Imatinib Mesylate; D010879:Piperazines; D011743:Pyrimidines; D051176:beta Catenin; D000099152:Cadherin 5; C495270:CTNNB1 protein, human