期刊文献+

苯丁酸钠通过上调p21WAF1/CIP1基因抑制白血病细胞系的细胞周期 被引量:3

Phenol butyrate inhibits cell cycle of leukemia cell lines through up-regulation of p21WAFI/CIP1 gene
原文传递
导出
摘要 目的研究组蛋白脱乙酰化酶(HDAC)抑制剂苯丁酸钠(PB)对白血病细胞系细胞周期的影响,探讨其分子机制。方法 PB 处理白血病细胞系 Kasumi-1、U937和 NB4细胞,分别于处理后24,48和72h 收集细胞。碘化丙锭 DNA 染色,流式细胞术分析细胞周期的变化。半定量逆转录-聚合酶链反应(RT-PCR)分析细胞周期相关基因 p21WAF1/CIP1表达的变化。在人肾上皮细胞系293T 细胞用荧光素酶报告基因分析 PB 对 p21WAF1/CIP1基因启动子活性的影响。结果 PB 可以抑制 Kasu-mi-1、U937和 NB4细胞的细胞周期,作用呈时间和剂量依赖关系。3 mmol/L PB 作用72 h,分别使 Ka-sumi-1、U937和 NB4细胞的 G_0/G_1期细胞比例增加42.03%、44.36%和26.82%,S 期细胞比例减少31.86%、38.91%和26.77%。PB 使 Kasumi-1、U937和 NB4细胞 p21WAF1/CIP1表达增高。PB 处理后,p21WAF1/CIP1的表达水平较处理前增高(2.06±0.27),(2.78±0.40)和(1.78±0.20)倍。PB 可以上调 p21 WAF1/CIP1启动子的转录活性,且呈剂量依赖关系。3 mmol/L PB 处理48 h 使转录活性增高(5.74±0.93)倍。PB 上调 p21WAF1/CIP1启动子转录活性主要是依赖于转录起始位点上游101 bp的序列。结论 PB 可以抑制白血病细胞系的细胞周期,这种作用可能是通过上调细胞周期相关基因p21 WAF1/CIP1的表达实现的。 Objective To elucidate effects of histone deacetylase inhibitors on cell cycle of leukemia cell lines and investigate its molecular mechanisms. Methods Kasumi-1, U937 and NB4 cell lines were exposed to a histone deacetylase inhibitor, phenyl butyrate(PB), for 24, 48 and 72 hrs. Cells were harvested for cell cycle analysis by flow cytometry. Gene expression of p21WAF1/CIP1, a cyclin-dependent kinase inhibitor, was determined by semi-quantitative reverse transcriptase polymerase chain reaction (semi-quantitative RT-PCR). Promoter activity of p21WAF1/CIP1 was determined by luciferase-reporter assay in 293T cell line. Results PB inhibited cell cycle of Kasumi-1, U937 and NB4 cell lines, showing G0/G1 phase arrest and S-phase fraction reduction with a dose and time dependent manner. After Kasumi-1, U937 or NB4 cells exposed to 3 mmoL/L PB for 72 hrs, G0/G1-phase fraction was increased by 42. 03% , 44. 36% and 26.82%, and S-phase fraction was decreased by 31.86%, 38.9%and 26.77%, respectively. After Kasumi-1, U937 and NB4 cell lines exposed to PB, the expression of p21WAF1/CIP1 gene was increased by (2.06±0.27), (2.78±0.40) and ( 1.78±0.20) times at its maximum, respectively. PB could stimulate p21WAF1/CIP1 promoter activity ( by luciferase-reporter assay) and the effect was dose dependent. The promoter activity was increased by 5.74 times after the cells exposed to 3 mmoL/L PB for 48 hrs. PB stimulating p21WAF1/CIP1 promoter activity was mainly mediated by a 101 base pairs fragment upstream of transcription start site. Conclusion PB could inhibit cell cycle of leukemia cell lines. The effects were mainly through up-regulation of p21WAF1/CIP1 expression.
出处 《中华血液学杂志》 CAS CSCD 北大核心 2006年第5期294-297,共4页 Chinese Journal of Hematology
基金 国家自然科学基金资助项目(30370593) 天津市科技发展计划资助项目(05YESZSF02400)
关键词 白血病 急性 抑制剂 组蛋白脱乙酰基酶 基因 p21WAF1/CIP1 Leukemia, acute Inhibitors, histone deacetylase Gene, p21WAF1/CIP1
  • 相关文献

参考文献6

  • 1郝长来,唐克晶,田征,邢海燕,王敏,王建祥.脱乙酰化酶抑制剂苯丁酸钠体外诱导Kasumi1细胞分化和凋亡作用[J].中华血液学杂志,2003,24(5):241-244. 被引量:13
  • 2el-Deiry WS,Tokino T,Velculescu VE,et al.WAF1,a potential mediator of p53 tumor suppression.Cell,1993,75:817-825.
  • 3Wang J,Hoshino T,Redner RL,et al.ETO,fusion partner in t (8;21) acute myeloid leukemia,represses transcription by interaction with the human N-CoR/mSin3/HDAC1 complex.Proc Natl Acad Sci U S A,1998,95:10860-10865.
  • 4Nakano K,Mizuno T,Sowa Y,et al.Butyrate activates the WAF1/Cip1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line.J Biol Chem,1997,272:22199-22206.
  • 5Lutterbach B,Westendorf JJ,Linggi B,et al.A mechanism of repression by acute myeloid leukemia-1,the target of multiple chromosomal translocations in acute leukemia.J Biol Chem,2000,275:651-656.
  • 6Suzuki A,Tsutomi Y,Akahane K,et al.Resistance to Fas-mediated apoptosis:activation of caspase 3 is regulated by cell cycle regulator p21 WAF1 and IAP gene family ILP.Oncogene,1998,17:931-939.

共引文献12

同被引文献25

引证文献3

二级引证文献3

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