ACSL3在前列腺癌细胞系中的表达及其对前列腺癌转移的影响

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英文篇名：Role of ACSL3 expression in suppressing prostate cancer cell metastasis 作者：李科 ; 陈怡 ; 董艳 ; 叶志强 英文作者：LI Ke;CHEN Yi;DONG Yan;YE Zhi-qiang;Department of Urology,The Third Affiliated Hospital of Sun Yat-sen University;Department of Breast Surgery,Sun Yat-sen Memorial Hospital of Sun Yat-sen University;Department of Biotechnology,School of Life Science and Technology,Jinan University;Department of Emergency,The Third Affiliated Hospital of Sun Yat-sen University; 关键词：前列腺肿瘤 ; 长链脂酰辅酶A合成酶3 ; 肿瘤侵袭 英文关键词：Prostate neoplasms;;Long-chain acyl-CoA synthetase 3;;Neoplasm invasiveness 中文刊名：ZBLS 英文刊名：Chinese Journal of Pathophysiology 机构：中山大学附属第三医院泌尿外科;中山大学孙逸仙纪念医院乳腺外科;暨南大学生命科学技术学院生物工程系;中山大学附属第三医院急诊科; 出版日期：2014-02-15 出版单位：中国病理生理杂志 年：2014 期：v.30 基金：广东省科技计划(No.2011B061300011) 语种：中文; 页：ZBLS201402012 页数：6 CN：02 ISSN：44-1187/R 分类号：62-67

摘要

目的:观察比较长链脂酰辅酶A合成酶3(long-chain acyl-CoA synthetase 3,ACSL3)在前列腺正常上皮细胞和不同类型前列腺癌细胞系中的表达差异,通过构建ACSL3基因稳定表达的前列腺癌细胞株研究ACSL3对前列腺癌细胞侵袭能力的影响。方法:利用RT-PCR方法检测ACSL3 mRNA在前列腺正常上皮细胞、局限性及转移性前列腺癌细胞中的表达情况,分析其表达与前列腺癌发生、进展及转移的关系;以前列腺癌细胞基因组cDNA为模板,通过PCR扩增ACSL3基因,重组构建含有Flag标签的pCDNA3.1(+)-Flag-ACSL3质粒,包装慢病毒,转染前列腺癌细胞,通过荧光显微镜、RT-PCR和Western blotting等方法鉴定ACSL3在细胞内的表达情况;利用Transwell侵袭实验研究ACSL3对前列腺癌细胞侵袭能力的改变。结果:较前列腺正常上皮细胞,各种前列腺癌细胞中ACSL3 mRNA表达均明显下降。同时,局限性前列腺癌细胞中ACLS3 mRNA表达较转移性前列腺癌细胞明显升高;此外,雄激素非依赖性前列腺癌细胞比雄激素依赖性前列腺癌细胞中ACLS3 mRNA表达显著降低。进而,我们成功构建和包装了表达ACSL3基因的pCDNA3.1(+)-Flag-ACSL3重组质粒及慢病毒,并转染前列腺癌细胞,筛选出稳定表达株;并且,通过Transwell实验证实ACSL3表达与前列腺癌细胞的侵袭能力呈负相关。结论:前列腺正常上皮细胞及不同前列腺癌细胞系中ACSL3的表达存在明显差异,提示ACSL3可能在前列腺癌的发生发展中起重要作用;成功构建了ACSL3基因稳定表达的前列腺癌细胞株,并初步证实ACSL3过表达可以抑制前列腺癌细胞的侵袭力。
        AIM:To analyze the difference of long-chain acyl-CoA synthetase 3(ACSL3) expression between normal prostate epithelial cells and prostate cancer cells.METHODS:ACLS3 mRNA expression between normal prostate epithelial cells and prostate cancer cells was compared using RT-PCR.Meanwhile,ACSL3 gene was amplified from prostate cancer cells,and the eukaryotic expression plasmid pCDNA3.1(+)-Flag-ACLS3 and lentivirus Lenti-ACLS3 were constructed.After transfection of ACSL3-plasmid and lentivirus into the prostate cancer cells,ACSL3 expressive was detected by RT-PCR and Western blotting,and then Matrigel invasion assay was performed to investigate the alteration of the invasive ability of the prostate cancer cells with over-expression of ACSL3.RESULTS:A significant difference of ACSL3 mRNA level between normal prostate epithelial cells and prostate cancer cells was observed.ACSL3 was highly expressed in localized prostate cancer cells compared to metastatic prostate cancer cells,while ACSL3 expression was higher in androgendependent prostate cancer cells than that in androgen-independent prostate cancer cells.Furthermore,the eukaryotic expression plasmid and lentivirus containing ACLS3 gene were successfully constructed.The prostate cancer cell line which stably over-expressed ACLS3 was established.Up-regulation of ACSL3 inhibited the invasive ability of prostate cancer cells.CONCLUSION:ACSL3 plays an antagonistic role in invasiveness of prostate cancer.

引文

[1]Siegel R,Naishadham D,Jemal A.Cancer statistics,2013[J].CA Cancer J Clin,2013,63(1):11-30.
    [2]Peyromaure M,DebréB,Mao K,et al.Management of prostate cancer in China:a multicenter report of 6 institutions[J].J Urol,2005,174(5):1794-1797.
    [3]Preston SH.Prostate-cancer screening[J].N Engl J Med,2009,361(2):202-203.
    [4]方习武,夏术阶,唐孝达.雄激素受体在人类前列腺癌发展中的作用研究[J].中国病理生理杂志,2004,20(7):1275-1279.
    [5]Marques RB,Dits NF,Erkens-Schulze S,et al.Modulation of androgen receptor signaling in hormonal therapy-resistant prostate cancer cell lines[J].PLoS One,2011,6(8):e23144.
    [6]Attard G,Glark J,Ambroisine L,et al.Heterogeneity and clinical significance of ETV1 translocations in human prostate cancer[J].Br J Cancer,2008,99(2):314-320.
    [7]Nchoutmboube JA,Viktorova EG,Scott AJ,et al.Increased long chain acyl-CoA synthetase activity and fatty acid import is linked to membrane synthesis for development of picornavirus replication organelles[J].PLoS Pathog,2013,9(6):e1003401.
    [8]Stamey TA,Caldwell M,McNeal JE,et al.The prostate specific antigen era in the United States is over for prostate cancer:what happened in the last 20 years?[J].J Urol,2004,172(4 Pt 1):1297-1301.
    [9]Bubendorf L,Schpfer A,Wagner U,et al.Metastatic patterns of prostate cancer:an autopsy study of 1,589 patients[J].Hum Pathol,2000,31(5):578-583.
    [10]王钢,王晓慧,张金山,等.雄激素受体基因(CAG)n重复多态性与前列腺癌[J].中国病理生理杂志,2000,16(10):1076.
    [11]Yao H,Ye J.Long chain acyl-CoA synthetase 3-mediated phosphatidylcholine synthesis is required for assembly of very low density lipoproteins in human hepatoma Huh7cells[J].J Biol Chem,2008,283(2):849-854.
    [12]Bu SY,Mashek MT,Mashek DG.Suppression of long chain acyl-CoA synthetase 3 decreases hepatic de novo fatty acid synthesis through decreased transcriptional activity[J].J Biol Chem,2009,284(44):30474-30483.
    [13]Cao A,Li H,Zhou Y,et al.Long chain acyl-CoA synthetase-3 is a molecular target for peroxisome proliferatoractivated receptor delta in HepG2 hepatoma cells[J].J Biol Chem,2010,285(22):16664-16674.
    [14]Mashek DG,Li LO,Coleman RA.Rat long-chain acylCoA synthetase mRNA,protein,and activity vary in tissue distribution and in response to diet[J].J Lipid Res,2006,47(9):2004-2010.
    [15]Fujimoto Y,Itabe H,Kinoshita T,et al.Involvement of ACSL in local synthesis of neutral lipids in cytoplasmic lipid droplets in human hepatocyte HuH7[J].Lipid Res,2007,48(6):1280-1292.