title: The expression and function of E3 ligase SIAH2 in acute T lymphoblastic leukemia.
abstract: BACKGROUND
  The seven in absentia homolog 2 (SIAH2) protein plays a significant role in human cancer by regulating hypoxia-inducible factor-a (HIF-1α); however, its role in T-cell acute lymphoblastic leukemia (T-ALL) is less clear.
  
  
  METHODS
  Immunofluorescence evaluation of SIAH2 protein expression and location were conducted in Jurkat cell (a T-ALL cell line) as well as in bone marrow mononuclear cells (BMMNCs) from T-ALL and idiopathic thrombocytopenic purpura (ITP) patients. The expression of SIAH2 mRNA was also examined by quantitative real-time PCR (qRT-PCR) in these cells. Lentivirus-packed shRNA targeting on SIAH2 (Lv-shSIAH2) was used to knock down SIAH2 expression in Jurkat cells. Cell proliferation, apoptosis, invasion and protein levels were then determined by CCK-8 assay, annexin V-PI assay, transwell and Western blotting, respectively.
  
  
  RESULTS
  The mRNA expression of SIAH2 in BMMNCs from primary T-ALL patients was significantly higher than cells from ITP patients (P=0.0312); There were significant positive associations between SIAH2 expression and the extramedullary infiltration (EMI) (P=0.0003), especially with the mediastinal lymph node metastasis (P=0.0168) and the pleural effusion (P=0.014). However, SIAH2 expression in T-ALL BMMNCs was not correlated with age, gender, white cell count or the clinical risk classification. SIAH2 knockdown by shRNA led to increased apoptosis and decreased proliferation, migration and invasion of Jurkat cells. Moreover, Prolyl Hydroxylase (PHD), P27 and Caspase3 were upregulated and HIF-1α, VEGF, VEGF Receptor 2, MMP-13, CyclinE1, C-myc and BCL2 were downregulated in SIAH2 knockdown Jurkat cells.
  
  
  CONCLUSIONS
  Our results suggest that SIAH2 regulates multi processes in T-ALL and may be an attractive therapeutic target.
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Hongxia
authors: H
authors: Chen
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Ning
authors: N
authors: Wang
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Guicun
authors: G
authors: Yang
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Yuxia
authors: Y
authors: Guo
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Yali
authors: Y
authors: Shen
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Xiaojing
authors: X
authors: Wang
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China.
authors: Ping
authors: P
authors: Zhang
authors: Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; Key Laboratory of Pediatrics in Chongqing, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China; China International Science and Technology Cooperation Center for Child Development and Disorders, Children's Hospital of Chongqing Medical University, Yuzhong, Chongqing 400014, PR China. Electronic address: feiyujidian@163.com.
authors: Youhua
authors: Y
authors: Xu
mesh_terms: D017209:Apoptosis / Q000235:genetics; D015153:Blotting, Western; D002465:Cell Movement / Q000235:genetics; D002648:Child; D005260:Female; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D006801:Humans; D019169:Jurkat Cells; D008297:Male; D009361:Neoplasm Invasiveness / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D000098563:Seven in Absentia Proteins
chemical_list: D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; D000098563:Seven in Absentia Proteins
keywords: HIF-1α; PHD; SIAH2; T-ALL