title: [Effects of DNMT1 Gene Silencing on Methylation of SOCS-1 Gene in Myeloma Cells].
abstract: OBJECTIVE
  To investigate the effect of DNA methyhransferase l (DNMT1) gene silencing on methylation of suppressor of cytokine signaling (SOCS-1) in multiple myeloma RPMI 8226 cells.
  
  
  METHODS
  Recombinant plasmid pshRNA-DNMTl was transfected into multiple myeloma RPMI 8226 cells by lipofectamine 2000. RT-PCR and Western blot were used to detect the mRNA and protein expression of DNMTl in RPMI 8226 cells respectively before and after transfection. Methylation-specific polymerase chain reaction was used to detect the methylation level change of SOCS-1 gene in RPMI8226 cells transfected.
  
  
  RESULTS
  DNMTl targeted short hairpin RNA(shRNA) was successfully inserted into the plasmid vector pshRNA. RT-PCR results showed that the relative mRNA expression level of DNMTI gene in RPMI 8226 cells transfected with pshRNA was 0.176±0.004 which was significantly lower than that in cells transfected by empty vector (0.956±0.033, P<0.01). Western blot analysis showed that the relative expression level of DNMT1 protein of RPMI 8226 cells transfected by pshRNA was 0.065±0.014, which was significantly lower than that in transfected cells by empty vector(0.415±0.027) (P<0.05). These results indicated that the recombinant plasmid pshRNA could effectively knock down the expression of DNMT1 gene in RPMI 8226 cells. Methylation analysis showed that the methylation level of SOCS-1 gene was obviously reduced after transfection.
  
  
  CONCLUSIONS
  DNMT1 gene in RPMI 8226 cell can be silenced by shRNA. DNMT1 gene silencing can significantly induce SOCS-1 gene hypomethylation, which indicates that DNMT1 may play an important role in the process of SOCS-1 hypermethylation.
authors.affiliation: Department of Hematology, Zhongshan Hospital Affiliated to Xiamen University; Fujian Medical University Teaching Hospital, Xiamen 361004, Fujian Province, China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Hematology, Zhongshan Hospital Affiliated to Xiamen University; Fujian Medical University Teaching Hospital, Xiamen 361004, Fujian Province, China. E-mail: nxq@xmu.edu.cn.
authors.forename: Xiao-Qing
authors.initials: XQ
authors.lastname: Niu
authors.affiliation: Department of Hematology, Zhongshan Hospital Affiliated to Xiamen University; Fujian Medical University Teaching Hospital, Xiamen 361004, Fujian Province, China.
authors.forename: Wen-Wen
authors.initials: WW
authors.lastname: Zhou
authors.affiliation: Department of Hematology, Zhongshan Hospital Affiliated to Xiamen University; Fujian Medical University Teaching Hospital, Xiamen 361004, Fujian Province, China.
authors.forename: Quan-Yi
authors.initials: QY
authors.lastname: Lu
mesh_terms: D045744:Cell Line, Tumor; D019175:DNA Methylation; D020868:Gene Silencing; D005822:Genetic Vectors; D006801:Humans; D009101:Multiple Myeloma; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D014162:Transfection
chemical_list: C411744:DMAP1 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C108944:SOCS1 protein, human; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins