唐卫军;张幸平;邹洪元;彭春芳;

• 1:重庆市江津区中心医院肿瘤科
• 2:重庆医科大学附属第一医院肿瘤科

摘要(Abstract)：

目的研究Skp2基因表达下调对HepG2细胞生物学特性及p27和SP1蛋白表达的影响。方法构建Skp2基因干扰慢病毒质粒,包装后感染HepG2细胞,48 h后,流式细胞仪检测细胞周期和凋亡情况;侵袭小室试验检测细胞侵袭能力;Western blot检测Skp2基因下调后p27和SP1蛋白的表达情况。结果与正常HepG2细胞组相比,Skp2干扰慢病毒组细胞Skp2蛋白的表达明显下调,p27蛋白表达大幅增加,细胞生长速度明显减慢,细胞凋亡率增加近两倍,G0/G1期细胞增多;SP1蛋白表达减少,浸润、迁移的细胞数减少约50%(P<0.05)。结论 Skp2蛋白表达下调抑制了HepG2细胞的增殖,促进HepG2细胞的凋亡,造成细胞侵袭能力减弱。Skp2蛋白表达下调导致p27蛋白表达上调、SP1蛋白表达下调,在上述生物学特性变化中可能发挥了重要作用,为深入探讨肝癌的病理机制奠定了基础。

关键词(KeyWords)： 肝肿瘤;Skp2基因;慢病毒载体;Sp1基因;p27基因

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 唐卫军;张幸平;邹洪元;彭春芳;

Email:

DOI: 10.13200/j.cjb.2012.01.23.tangwj.028

参考文献(References)：

• [1]中国抗癌协会肝癌专业委员会,中国抗癌协会临床肿瘤学协作委员会,中华医学会肝病学分会肝癌学组,等.原发性肝癌规范化诊治专家共识[J].临床肿瘤学杂志,2009,14(3):259-269.
• [2]Fasanaro P,Capogrossi MC,Martelli F.Regulation of the en-dothelial cell cycle by the ubiquitin-proteasome system[J].Car-diovasc Res,2010,85(2):272-280.
• [3]Frescas D,Pagano M.Deregulated proteolysis by the F-box pro-teins SKP2and beta-TrCP:tipping the scales of cancer[J].Nat Rev Cancer,2008,8(6):438-449.
• [4]Zhu L.Skp2knockout reduces cell proliferation and mouse body size:and prevents cancer[J].Cell Res,2010,20(6):605-607.
• [5]Su Y,Wang F,Qi H,et al.Small interfering RNA targeting of S phase kinase-interacting protein2inhibits cell proliferation of pterygium fibroblasts[J].Mol Vis,2011,17(1):247-256.
• [6]Tan NY,Khachigian LM.Sp1phosphorylation and its regulation of gene transcription[J].Mol Cell Biol,2009,29(10):2483-2488.
• [7]Deniaud E,Baguet J,Chalard R,et al.Overexpression of tran-scription factor Sp1leads to gene expression perturbations and cell cycle inhibition[J].PLoS One,2009,4(9):e7035.
• [8]Yuan P,Wang L,Wei D,et al.Therapeutic inhibition of Sp1ex-pression in growing tumors by mithramycin a correlates directly with potent antiangiogenic effects on human pancreatic cancer[J].Cancer,2007,110(12):2682-2690.
• [9]王丽杰,史晓宇,周建华.转录因子Sp1及其与肿瘤的关系[J].实用肿瘤学杂志,2008,22(5):478-480.
• [10]白雪,邓红.转录因子Sp1与肿瘤关系研究的新进展[J].浙江大学学报(医学版),2010,39(2):215-220.
• [11]Tapias A,Ciudad CJ,Roninson IB,et al.Regulation of Sp1by cell cycle related proteins[J].Cell Cycle,2008,7(18):2856-2867.
• [12]Hung WC,Tseng WL,Shiea J,et al.Skp2overexpression in-creases the expression of MMP-2and MMP-9and invasion of lung cancer cells[J].Cancer Lett,2010,288(2):156-161.
• [13]Takanami I.The prognostic value of overexpression of Skp2mR-NA in non-small cell lung cancer[J].Oncol Rep,2005,13(4):727-73l.
• [14]罗欣,刘丹丹,漆洪波,等.靶向人Gadd45α基因的shRNA慢病毒载体对缺氧致人脐静脉内皮细胞生物学功能损伤的影响[J].中国生物制品学杂志,2010,23(11):1168-1172.
• [15]房丽君,黄世威,王颖,等.狂犬病病毒糖蛋白基因重组慢病毒载体的构建及鉴定[J].中国生物制品学杂志,2011,24(2):130-132.
• [16]陈国庆,姚珍薇,秦兴发.靶向人转录因子FOXM1基因的shRNA慢病毒载体的构建与鉴定[J].中国生物制品学杂志,2011,24(9):1050-1053.
• [17]Sutterlüty H,Chatelain E,Marti A,et al.p45SKP2promotes p27Kip1degradation and induces S phase in quiescent cells[J].Nat Cell Biol,1999,1(4):207-214.
• [18]Carrano AC,Eytan E,Hershko A,et al.SKP2is required for ubiquitin-mediated degradation of the CDK inhibitor p27[J].Nat Cell Biol,1999,1(4):193-199.
• [19]Sumimoto H,Yamagata S,Shimizu A,et al.Gene therapy for human small-cell lung carcinoma by inactivation of Skp2with virally mesiated RNA interference[J].Gene Ther,2005,12(1):95-100.
• [20]Ezoe S,Matsumura I,Nakata S,et al.GATA-2/estrogen receptor chimera regulates cytokine-dependent growth of hematopoietic cells through accumulation of p21 WAFI and p27 Kip1 protei ns[J].Blood,2002,100(10):3512-3520.
• [21]Cen B,Deguchi A,Weinstein IB.Activation of protein kinase G increases the expression of p21 CIP1,p27 KIP1,and histidine triad protein1through Sp1[J].Cancer Res,2008,68(13):5355-5362.