title: [Abnormal WT1 gene expression in paroxysmal nocturnal hemoglobinuria].
abstract: OBJECTIVE
  To explore the pathogenesis of abnormal WT1 expression in paroxysmal nocturnal hemoglobinuria (PNH).
  
  
  METHODS
  The expression of WT1 mRNA in CD59⁻ and CD59⁺ bone marrow mononuclear cells (BMMNC) were measured by semi-quantitative reverse transcription PCR. After WT1 gene silence by RNA interference (RNAi) technology, biological characteristics of BMMNC were investigated by flow cytometry.
  
  
  RESULTS
  The relative expression of WT1 mRNA in PNH CD59⁻ BMMNC (1.06 ± 0.12) was significantly higher than that in PNH CD59⁺ BMMNC (0.90 ± 0.12) and normal BMMNC (0.86 ± 0.05, P<0.05), but there was no significant difference between PNH CD59⁺ BMMNC and normal BMMNC (P>0.05). WT1 mRNA expression in PNH was positively correlated with the proportion of CD59⁻ cells (r²=0.490, P=0.016), but had no relationship with the proportion of CD59⁺ cells. After WT1 gene silence by siRNA in PNH CD59⁻ BMMNC, WT1 mRNA expression was decreased. The proportions of G0/G1 phase in PNH CD59⁻ cell blank control group and siRNA-scr transfected group were (92.73 ± 3.71)% and (93.06 ± 4.14)%, and the proportions of S phase were (6.99 ± 3.61)% and (6.73 ± 4.08)%, respectively. The proportions of G0/G1 and S phase in siRNA-WT1 transfected group was (94.46 ± 3.71)% and (5.40 ± 3.55)%, respectively. There were significant differences in the proportions of G0/G1 phase and S phase among the controls, siRNA-WT1 transfected group and siRNA-scr transfected group (P<0.05). The rate of apoptosis in siRNA-WT1 transfected group [(35.91 ± 22.36)%] was significantly higher than those in controls [(26.12 ± 17.10)%] and siRNA-scr transfected group [(27.39 ± 18.99)%] (P<0.05).
  
  
  CONCLUSIONS
  siRNA-WT1 could effectively suppress the WT1 gene expression of CD59⁻ clone in PNH patients, inhibit its proliferation, and promote its apoptosis. WT1 gene expression might contribute to PNH clone proliferation.
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Yuanyuan
authors: Y
authors: Zhang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Rong
authors: R
authors: Fu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Yihao
authors: Y
authors: Wang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Lijuan
authors: L
authors: Li
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Hui
authors: H
authors: Liu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Chunyan
authors: C
authors: Liu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Tian
authors: T
authors: Zhang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Shaoxue
authors: S
authors: Ding
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Liyan
authors: L
authors: Li
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Erbao
authors: E
authors: Ruan
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Wen
authors: W
authors: Qu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Huaquan
authors: H
authors: Wang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Xiaoming
authors: X
authors: Wang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Guojin
authors: G
authors: Wang
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Yuhong
authors: Y
authors: Wu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Jia
authors: J
authors: Song
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Hong
authors: H
authors: Liu
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Limin
authors: L
authors: Xing
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Jing
authors: J
authors: Guan
authors: Department of Hematology, General Hospital, Tianjin Medical University, Tianjin 300052, China.
authors: Zonghong
authors: Z
authors: Shao
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000378:metabolism; D002453:Cell Cycle; D005260:Female; D006457:Hemoglobinuria, Paroxysmal / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D025721:WT1 Proteins / Q000235:genetics / Q000378:metabolism*; D055815:Young Adult
chemical_list: D012333:RNA, Messenger; D025721:WT1 Proteins