期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

沉默DOT1L基因对急性单核细胞白血病细胞THP-1增殖的影响 被引量：3

Effect of DOT1LGene Silence on Proliferation of Acute Monocytic Leukemia Cell Line THP-1
暂未订购
导出
摘要 本研究旨在探讨DOT1L基因对人白血病THP-1细胞增殖的影响。针对DOT1L mRNA的第732-752靶位点设计并合成shRNA,构建pLKO-Tet-On条件性干扰载体,制备慢病毒并感染THP-1细胞,用强力霉素(Dox)诱导干扰载体表达;应用RT-PCR的方法验证干扰效率;采用改良MTT法检测干扰DOT1L基因后对THP-1细胞增殖能力的影响;用细胞集落形成实验观察干扰后细胞集落形成的能力;用流式细胞术分析细胞周期的变化。结果表明:THP-1细胞干扰后DOT1L基因的表达显著降低;DOT1L基因的表达下调抑制了细胞的增殖能力、集落形成能力并将细胞周期阻滞在G0/G1期。结论:利用shRNA靶向干扰DOT1L基因的表达,可以有效抑制人急性单核细胞白血病细胞THP-1的增殖。 This study was aimed to investigate the influence of short hairpin RNA（shRNA） on proliferation of human leukemia cell line THP-1.The shRNA targeting the site 732-752 of DOT1L mRNA was designed and chemically synthesized,then a single-vector lentiviral,tet-inducible shRNA-DOT1L system（Plko-Tet-On） was generated.Thereafter,the THP-1 cells with lentivirus were infected to create stable cell line with regulatable shRNA expression.The expression of DOT1L in the THP-1 cell line was assayed by RT-PCR.Effect of shRNA-DOT1L on the proliferation of THP-1 cells was detected with MTT method,and the change of colony forming potential of THP-1 cells was analyzed by colony forming unit test.Cell cycle distribution was tested by flow cytometry.The results indicated that the expression of DOT1L was statistically lower than that in the control groups.The proliferation and colony forming capacity of THP-1 cells were significantly inhibited.The percentage of cells at G0/G1 phase increased in THP-1/shRNA cells treated with Dox while the percentage of cells at S phase significantly decreased as compared with that in the control group.It is concluded that the shRNA targeting DOT1L can effectively inhibit the proliferation of acute monocytic leukemia cell line THP-1.
作者 张玉娟 李华文 常国强 张洪菊 王建 蔺亚妮 周家喜 李庆华 庞天翔
机构地区 中国医学科学院、北京协和医学院血液学研究所、血液病医院
出处 《中国实验血液学杂志》 CAS CSCD 北大核心 2013年第4期861-865,共5页 Journal of Experimental Hematology
关键词 DOT1L基因 急性单核细胞白血病 THP-1细胞 DOT1L gene acute monocytic leukemia THP-1 cell
分类号 R733.71 [医药卫生—肿瘤]
  • 相关文献

参考文献14

  • 1Ernst P, Wang J, Korsmeyer SJ. The role of MLL in hematopoiesis and leukemia. Curr Opin Hematol, 2002 ; 9 (4) : 282 - 287.
  • 2Canaani E, Nakamura T, Rozovskaia T, et al. ALL-1/MLL1, a homologue of Drosophila TRITHORAX, modifies chromatin and is di- reetlyinvolved in infant acute leukaemia. Br J Cancer, 2004; 90 (4) : 756 -760.
  • 3Krivtsov AV, Armstrong SA. MLL translocations, histone modifica- tions and leukaemia stem-cell development. Nat Rev Cancer, 2007; 7(11) : 823 -833.
  • 4Dou Y, Hess JL. Mechanisms of transcriptional regulation by MLL and its disruption in acute leukemia. Int J Hematol, 2008 ; 87 ( 1 ) : 10 - 18.
  • 5Liedtke M, Cleary ML. Therapeutic targeting of MLL. Blood, 2009 ; 113(24) : 6061 -6068.
  • 6Nguyen AT, Zhang Y. The diverse functions of Dotl and H3K79 methylation. Genes Dev, 2011 ; 25(13) : 1345 - 1358.
  • 7Ng HH, Feng Q, Wang H, et al. Lysine methylation within the globular domain of histone H3 by Dotl is important for telomerie si- lencingand Sir protein association. Genes Dev, 2002; 16 ( 12 ) : 1518 - 1527.
  • 8van Leeuwen F, Gafken PR, Gottsehling DE. Dotlp modulates si- fencing in yeast by methylation of the nueleosome core. Cell, 2002 ; 109(6) : 745 -756.
  • 9Feng Q, Wang H, Ng HH, et al. Methylation of H3-1ysine 79 is me- diated by a new family of HMTases without a SET domain. Curr Bi- al, 2002; 12(12) : 1052 - 1058.
  • 10Lacoste N, Utley RT, Hunter JM, et al. Disruptor of telomeric silen- cing-1 is a chromatin-specific H3 methyl-transferase. J Biol Chem, 2002 ; 277 (34) : 30421 - 30424.

二级参考文献7

  • 1Xia ZB, Popovic R, Chen J, et al. The MLL fusion gene, MLLAF4, regulates cyclin-dependent kinase inhibitor CDKN1B ( p27kipl ) expression. Proc Natl Acad Sci USA, 2005 ; 102 : 14028 - 14033
  • 2Hammond SM, Caudy AA, Hannon GJ. Post-transcriptional gene silencing by double-stranded RNA. Nat Rev Genet,2001 ; 2 : 110 - 119
  • 3Odero MD, Zeleznik-Le NJ, Chinwalla V, et al. Cytogenetic and molecular analysis of the acute monocytic leukemia cell line THP-1 with an mll-af9 translocation. Genes Chromosomes Cancer, 2000; 29:333 - 338
  • 4Li ZY, Liu DP, Liang CC. New insight into the molecular mechanisms of MLL-associated leukemia. Leukemia, 2005 ; 19 : 183 - 190
  • 5Shin S, Sung B J, Cho YS, et al. An anti-apoptotic protein human survivin is a direct inhibitor of easpase-3 and-7. Biochemistry, 2001 ; 40:1117 -1123
  • 6Kawagoe H, Kawagoe R, Sano K. Targeted down-regulation of mll-af9 with antisense oligodeoxyribonucleotide reduces the expression of the HOXA7 and -A10 genes and induces apoptosis in a human leukemia cell line, THP-1. Leukemia, 2001 ; 15 : 1743 - 1749
  • 7Pession A, Martino V, Tonelli R, et al. mll-af9 oncogene expression affects cell growth but not terminal differentiation and is downregulated during monocyte-macrophage maturation in AMLM5 THP-1 cells. Oncogene, 2003 ;22:8671 - 8676

共引文献1

同被引文献27

  • 1李庆华,庞天翔,韩忠朝.敲除Sam68基因导致白血病细胞系细胞生长迟缓和G2/M期延长[J].生物化学与生物物理进展,2005,32(8):781-787. 被引量：2
  • 2Lukong KE,Richard S.Targeting the RNA-binding protein Sam68as a treatment for cancer?Future Oncol,2007;3(5):539-544.
  • 3Rajan P,Gaughan L,Dalgliesh C,et al.The RNA-binding and adaptor protein Sam68 modulates signal-dependent splicing and transcriptional activity of the androgen receptor.J Pathol,2008;215(1):67-77.
  • 4Brignatz C,Paronetto MP,Opi S,et al.Alternative splicing modulates autoinhibition and SH3 accessibility in the Src kinase Fyn.Mol Cell Biol,2009;29(24):6438-6448.
  • 5Chawla G,Lin CH,Han A,et al.Sam68 regulates a set of alternatively spliced exons during neurogenesis.Mol Cell Biol,2009;29(1):201-13.
  • 6Babic I,Cherry E,Fujita DJ.SUMO modification of Sam68enhances its ability to repress cyclin D1 expression and inhibits its ability to induce apoptosis.Oncogene,2006;25(36):4955-4964.
  • 7Busa R,Paronetto MP,Farini D,et al.The RNA-binding protein Sam68 contributes to proliferation and survival of human prostate cancer cells.Oncogene,2007;26(30):4372-4382.
  • 8Richard S,Vogel G,Huot ME,et al.Sam68 haploinsufficiency delays onset of mammary tumorigenesis and metastasis.Oncogene,2008;27(4):548-556.
  • 9Elliott DJ,Rajan P.The role of the RNA-binding protein Sam68 in mammary tumourigenesis.J Pathol,2010;222(3):223-226.
  • 10Zhang Z,Li J,Zheng H,et al.Expression and cytoplasmic localization of Sam68 is a significant and independent prognostic marker for renal cell carcinoma.Can Epid Biomark Prev,2009;18(10):2685-2693.

引证文献3

二级引证文献5

中国实验血液学杂志
2013年 第4期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部