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SUMO-1基因siRNA抑制肝癌细胞SMMC-7721生长的研究 被引量：3

The Growth Inhibition Induced by SUMO-1 siRNA in Hepatocellular Carcinoma Cell Line SMMC-7721
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摘要 目的:研究SUMO-1基因siRNA沉默人肝癌细胞SMMC-7721中SUMO-1基因表达的效果及对SMMC-7721生长的影响。方法:将人工合成的针对SUMO-1基因的siRNA片段转染体外培养的人肝癌细胞SMMC-7721,采用RT-PCR、Westernblot方法在mRNA及蛋白水平检测siRNA沉默肝癌细胞中SUMO-1基因的效果。通过MTT、流式细胞仪及TUNEL试验检测SUMO-1基因沉默后对肝癌细胞生长的影响。结果:人工合成的针对SUMO-1基因的siRNA片段能显著地抑制SMMC-7721中SUMO-1基因的表达,48h抑制率可达73.43%。MTT结果显示肝癌细胞SMMC-7721转染SUMO-1siRNA后生长明显受到抑制,流式细胞仪检测G2期细胞明显增加,但TUNEL试验未发现凋亡细胞。结论:SUMO-1siRNA沉默肝癌细胞SMMC-7721中SUMO-1基因效果良好,SUMO-1基因控制SMMC-7721的生长,SUMO-1基因是肝癌生长的一个重要的调控因素。 Objective：To study the efficiency of silencing small ubiquitin-like modifier-1 （SUMO-1） induced by siRNA in hepatocellular carcinoma cell line SMMC-7721 and the growth inhibition of SMMC-7721 thereof. Methods：The SUMO-1 siRNA was transfected into SMMC-7721 by means of lipofectamineTM 2000. The silencing efficiency of SUMO-1 was examined by RT-PCR and western blot. The cell growth and cell cycle were examined by MTT and flow cytometry （FCM）. The cell apoptosis was detected by DeadEndTM Colorimetric TUNEL System. Results：The siRNA could significantly silence the expression of SUMO-1 in SMMC-7721.The maximal silencing rate was utmost 73.43% at 48 hours after being transfected SUMO-1 siRNA. MTT assay revealed that the cell line grew more slowly. FCM result showed that the number of G2 stage cells was increased significantly. But apoptosis cells were not found by TUNNEL assay. Conclusion：SiRNA is a good manner to silence the expression of SUMO-1 in SMMC-7721 in vitro. Owing to the growth inhibition induced by SUMO-1 siRNA,SUMO-1 plays an important role in development of SMMC-7721.
作者 郭武华 袁丽华 肖志华 罗良平 张吉翔
机构地区 南昌大学第二附属医院肿瘤科 江西省分子生物重点实验室 南昌大学第二附属医院消化科
出处 《天津医药》 CAS 北大核心 2010年第1期4-6,81,共4页 Tianjin Medical Journal
基金 江西省教育厅科技支撑项目(项目编号:GJJ09107)
关键词 肝肿瘤 癌 细胞系 肿瘤 RNA 小分子干扰 基因沉默 liver neoplasms carcinoma cell line tumor RNA small interfering gene silencing
分类号 R285.5 [医药卫生—中药学] R735.7 [医药卫生—肿瘤]
  • 相关文献

参考文献7

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二级参考文献19

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共引文献6

同被引文献26

  • 1Di Bacco A,Ouyang J,Lee HY,et al.The SUMO-specific protease SENP5 is required for cell di-vision[J].Mol Cell Biol,2006,26(12):4489.
  • 2Martin S,Nishimune A,Mellor JR,et al.SUMO ylation regulates kainate-receptor-mediated synaptic transmission[J].Nature,2007,447(7142):321.
  • 3Buschmann T,Fuchs SY,Lee CG,et al.SUMO-1 modification of Mdm2 prevents its self-ubiquitination and increases Mdm2 ability to ubiquitinate p53[J].Cell,2000,101(7):753.
  • 4Carter S,Bischof O,Dejean A,et al.C-terminal modifications regulate MDM2 dissociation and nuclear export of p53[J].Nat Cell Biol,2007,9(4):428.
  • 5Pfander B,Moldovan GL,Sacher M,et al.SUMO-modified PCNA recruits Srs2 to prevent recombination during S phase[J].Nature,2005,436(7049):428.
  • 6Karamouzis MV,Konstantinopoulos PA,Badra FA,et al.SUMO and estrogen receptors in breast cancer[J].Breast Cancer Res Treat,2008,107(2):195.
  • 7Xhemalce B,Riising EM,Baumann P,et al.Role of SUMO in the dynamics of telomere maintenance in fission yeast[J].Proc Natl Acad Sci USA,2007,104(3):893.
  • 8Ulrich HD.Mutual interactions between the SUMO and ubiquitin systems:a plea of no contest[J].Trends Cell Biol,2005,15(10):525.
  • 9Lomeli H, Vazquez M. Emerging roles of the SUMO pathway in development[J]. Cell Mol Life Sci, 2011,68 (24) :4045-4064.
  • 10Ulrich HD. Ubiquitin,SUMO,and Phosphate: How a trio of posttranslational modifiers governs protein fate [J]. Mol Ce11,2012,47(3) :335-337.

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天津医药
2010年 第1期

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