贺珊,吴维光,王筝,闫洪亮,孙兆翎,唐雅娟
HE Shan,WU Weiguan,WANG Zheng,YAN Hongliang,SUN Zhaoling,TANG Yajuan

• 1:武警后勤学院附院妇产科

摘要(Abstract)：

目的:观察组蛋白去乙酰化酶1(HDAC1)基因沉默对人卵巢癌SKOV3细胞迁移及侵袭能力的影响。方法:体外合成针对HDAC1的小干扰RNA(siRNA),利用脂质体转染入人卵巢癌SKOV3细胞;通过Western blot方法检测细胞内HDAC1蛋白和乙酰化组蛋白H4(Ac-H4)的表达,利用荧光定量PCR方法检测HDAC1、尿激酶型纤溶酶原激活因子(uPA)和尿激酶型纤溶酶原激活因子受体(uPAR)基因的mRNA表达;通过划痕实验检测细胞迁移能力,利用Transwell侵袭实验检测细胞侵袭能力。结果:经转染HDAC1基因siRNA后,卵巢癌SKOV3细胞HDAC1基因mRNA和蛋白表达水平均下调,细胞内组蛋白H4乙酰化水平增加;SKOV3细胞迁移速度降低,穿过Transwell滤膜的细胞数量减少;SKOV3细胞内uPA和uPAR基因mRNA表达也降低。结论:HDAC1基因沉默能够抑制人卵巢癌SKOV3细胞迁移和侵袭能力,机制可能与增加组蛋白乙酰化水平、抑制uPA和uPAR基因表达有关。
Objective: To observe the effect of histone deacetylase 1( HDAC1) gene silence on migration and invasion of human ovarian cancer cell line SKOV3. Methods: The small interference RNA( siRNA) targeting HDAC1 was transfected into human ovarian cancer SKOV3 cells by lipofectamine2000. Western blot was used to detect the HDAC1 protein and level of acetyl histone H4,and real time PCR was used to detect the mRNA levels of HDAC1,urokinase plasminogen activator( uPA) and urokinase plasminogen activator receptor( uPAR) genes. Furthermore,cell migration was estimated by wound-healing assay and cell invasion was estimated by Transwell assay. Results: After transfection,the levels of HDAC1 of ovarian cancer cell line SKOV3 mRNA and protein were decreased,and the acetyl level of histone H4 was increased. The cell migration and invasion number of cells were decreased. Meanwhile,the expressions of uPA and uPAR mRNA were decreased in SKOV3 cells. Conclusions: The siRNA targeting HDAC1 may inhibit the migration and invasion activity of ovarian cancer SKOV3 cells,the mechanism is possible to down-regulate uPA and uPAR gene expressions by increasing acetyl level of histone.

关键词(KeyWords)： 卵巢;癌;组蛋白脱乙酰化酶类;基因沉默;细胞迁移;细胞侵袭
ovarian cancer;histone deacetylases;gene silencing;cell migration;cell invasion

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 贺珊,吴维光,王筝,闫洪亮,孙兆翎,唐雅娟
HE Shan,WU Weiguan,WANG Zheng,YAN Hongliang,SUN Zhaoling,TANG Yajuan

DOI: 10.19367/j.cnki.1000-2707.2014.05.025

参考文献(References)：

• [1]Liu S.Epigenetices advancing personalized nanomedicine in cancer therapy[J].Adv Drug Deliv Rev,2012(13):1532-1543.
• [2]Fang Y,Hu Y,Wu P,et al.Synergistic efficacy in human ovarian cancer cells by histone deacethylase inhibitor TSA and proteasome inhibitor NS-341[J].Cancer Invest,2011(4):247-252.
• [3]Jeon HS,Ahn MY,Park JH,et al.Anticancer effects of the MHY218 novel hydroxamic acid-derived histone deacetylase inhibitor in human ovarian cancer cells[J].Int J Oncol,2010(2):419-428.
• [4]Hwang JJ,Kim YS,Kim MJ,et al.A novel histone deacetylase inhibitor,CG006,induces cell death through both extrinsic and intrinsic apoptotic pathways[J].Anticancer Drugs,2009(9):815-821.
• [5]Hagelkruys A,Sawicka A,Selser C,et al.The biology of HDAC in cancer;the nuclear and epigenetic components[J].Handb Exp Pharmacol,2011(206):13-37.
• [6]贾妍,邹颖刚,陈军,等.组蛋白去乙酰化酶1在人上皮性卵巢癌组织中的表达[J].中国实验诊断学杂志,2011(2):256-258.
• [7]高道键,徐岷,张玉琦,等.RNA干扰组蛋白去乙酰化酶1对人胰腺癌细胞增殖、凋亡的调控机制研究[J].解放军医学杂志,2010(2):380-383.
• [8]李娜,于世英.组蛋白去乙酰化酶1siRNA对HeLa细胞生长和凋亡的影响[J].华中科技大学学报:医学版,2010(1):47-49.
• [9]顾红军,董宇超,李强.HDAC1 siRNA转染对肺腺癌A549细胞增殖及凋亡的影响[J].中国癌症杂志,2010(8):578-582.
• [10]潘其壮,覃展偶.uPA系统与肿瘤的研究进展[J].医学综述杂志,2010(14):2140-2143.
• [11]AI-Hassan NN,Behzadlau A,Caldwell P,et al.Differential roles of uPAR in peritoneal ovarian carcinomatosis[J].Neoplasia,2012(4):259-270.
• [12]许天敏,崔满华,林杨,等.uPA、uPAR和PAI-1在恶性卵巢上皮性肿瘤中的表达及意义[J].中国实验诊断学杂志,2009(10):1371-1372.

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