JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES)

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Changes of NF-κB expression after promoter demethylation of TMS1 in K562 cells

LI Hongli, WANG Yan, XU Wenwei, DONG Lin, GUO Yan, ZHU Chuansheng, BI Kehong   

  1. Department of Hematology, Qianfoshan Hospital Affiliated to Shandong University, Jinan 250014, Shandong, China
  • Received:2013-10-22 Online:2014-04-10 Published:2014-04-10

Abstract: Objective  To explore the association between the methylation degree of TMS1 gene in chronic myeloid leukemia cell line K562 cell and NF-κB. Methods  A total of 6 samples of bone marrow of healthy controls and patients with non-hematologic malignancies were randomly collected as case-control group. Then total RNA was extracted and TMS1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). K562 cells were cultured and treated with decitabine (DCA) with different concentrations. The blank control group and experiment group (0.5μmol/L DCA, 1μmol/L DCA) were set up separately. Methylated degree of TMS1 was detected by methylation specificity polymerase chain reaction (MS-PCR), mRNA and protein of TMS1 and NF-κB in K562 cells were checked by RT-PCR and Western blot, the growth of cells was inspected by CCK-8 and morphologic changes were observed by light microscope. Results  TMS1 mRNA was found in the case-control group, but was absent in K562 cells. The TMS1 gene was exhaustively methylated in the blank control group. After promoter demethylation of TMS1, the expression of NF-κB significantly decreased (P<0.05). The K562 cell proliferation was inhibited in a dose- and time-dependent manner. Conclusion  TMS1 gene presents exhaustive methylation in K562 cells and its silent expression may abnormally activate NF-κB signaling pathway and plays a role in the pathogenesis of leukemia.

Key words: Leukemia, Methylation, Gene, TMS1 gene; 

CLC Number: 

  • R733.7
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