高级搜索

沉默SEPT9基因对肝癌HepG2细胞增殖及凋亡的影响

曾永秋, 曹洋, 梅志强, 刘岚, 税青林

曾永秋, 曹洋, 梅志强, 刘岚, 税青林. 沉默SEPT9基因对肝癌HepG2细胞增殖及凋亡的影响[J]. 肿瘤防治研究, 2013, 40(03): 236-239. DOI: 10.3971/j.issn.1000-8578.2013.03.004
引用本文: 曾永秋, 曹洋, 梅志强, 刘岚, 税青林. 沉默SEPT9基因对肝癌HepG2细胞增殖及凋亡的影响[J]. 肿瘤防治研究, 2013, 40(03): 236-239. DOI: 10.3971/j.issn.1000-8578.2013.03.004
Zeng Yongqiu, Cao Yang, Mei Zhiqiang, Liu Lan, Shui Qinglin. Effects of SEPT9 Gene Silencing on Cell Proliferation and Apoptosis of HepG2 Hepatoma Cells[J]. Cancer Research on Prevention and Treatment, 2013, 40(03): 236-239. DOI: 10.3971/j.issn.1000-8578.2013.03.004
Citation: Zeng Yongqiu, Cao Yang, Mei Zhiqiang, Liu Lan, Shui Qinglin. Effects of SEPT9 Gene Silencing on Cell Proliferation and Apoptosis of HepG2 Hepatoma Cells[J]. Cancer Research on Prevention and Treatment, 2013, 40(03): 236-239. DOI: 10.3971/j.issn.1000-8578.2013.03.004

沉默SEPT9基因对肝癌HepG2细胞增殖及凋亡的影响

基金项目: 四川省卫生厅基金资助项目(100211)
详细信息
    作者简介:

    曾永秋(1980-),女,硕士,讲师,主要从事肿瘤分子遗传学的研究

    通讯作者:

    税青林,E-mail:shuiql889@163.com

  • 中图分类号: R394.3;R735.7

Effects of SEPT9 Gene Silencing on Cell Proliferation and Apoptosis of HepG2 Hepatoma Cells

More Information
    Corresponding author:

    Shui Qinglin, E-mail:shuiql889@163.com

  • 摘要: 目的 探讨SEPT9基因与肝癌的关系,进一步揭示SEPT9基因在肝癌发生发展过程中的作用。方法将构建的针对SEPT9基因的shRNA表达载体通过脂质体LipofectamineTM2000转染人肝癌HepG2细胞,荧光显微镜下观察细胞的转染效率,RT -PCR、Western blot法分别检测SEPT9 mRNA和蛋白质的表达抑制情况,CCK-8检测细胞的生长抑制情况,流式细胞术检测细胞凋亡情况。结果荧光显微镜观察细胞的转染效率达到70%以上;SEPT9 mRNA及蛋白表达抑制率均受到明显抑制。CCK-8法检测结果显示,实验组HepG2细胞的生长受到明显抑制;流式细胞术检测结果显示实验组HepG2细胞凋亡明显(P<0.05)。结论抑制SEPT9基因的表达可有效抑制HepG2细胞的增殖,并对细胞凋亡有明显的促进作用。

     

    Abstract: Objective To explore the relationship between SEPT9 gene and hepatocellular carcinoma,and to reveal the role of SEPT9 in tumorigenesis and development of hepatoma. Methods The expression vector containing SEPT9 gene was transfected into the human hepatoma cell line HepG2 with LipofectamineTM2000.The transfection efficacy was studied under fluorescence microscopy,and the mRNA and protein levels of SEPT9 were detected with reverse transcript polymerase chain reaction (RT-PCR) and Western blot assay.CCK-8 method was used to detect cell proliferation,and FCM was served to detect the cell apoptosis. Results The transfection efficiency was up to 70%,and expression of mRNA and protein of SEPT9 were suppressed significantly.And the result of CCK-8 assay showed significantly inhibition of in the experiment group,and FCM assay showed that the apoptotic rate in the experiment group increased significantly comparing to the control group (P<0.05 ). Conclusion Suppression of the expression of SEPT9 gene can effectively inhibit the proliferation of HepG2 cell,and promote apoptosis significantly.

     

  • [1] Dobbelaere J,Barral Y.Spatial coordination of cytokinetic events by compartmentalization of the cell cortex[J].Science,2004,305(5682):393-6.
    [2] Spiliotis ET,Kinoshita M,Nelson WJ.A mitotic septin scaffold required for mammalian chromosome congression and segregation[J].Science,2005,307(5716):1781-5.
    [3] Kartmann B,Roth D.Novel roles for mammalian septins:from vesicle trafficking to oncogenesis[J ].J Cell Sci,2001,114(Pt 5):839-44.
    [4] Gottfried Y,Rotem A,Lotan R,et al.The mitochondrial ARTS protein promotes apoptosis through targeting XIAP[J].EMBO J,2004,23(7):1627-35.
    [5] Hall PA,Russell SH.The pathobiology of the septin gene family[J].J Pathol,2004,204(4):489- 505.
    [6] Larisch S.The ARTS connection:role of ARTS in apoptosis and cancer[J].Cell Cycle,2004,3(8): 1021-3.
    [7] Ihara M,Tomimoto H,Kitayama H,et al.Association of the cytoskeletal GTP-binding protein Sept4/H5 with cytoplasmic inclusions found in Parkinson’s disease and other synucleinopathies[J].J Biol Chem,2003,278(26):24095-102.
    [8] Cossart P,Pizarro-Cerda J,Lecruit M.Invasion of mammalian cells by Listeria monocytogenes: functional mimicry to subvert cellular functions[J].Trends Cell Biol,2003,13(1):23-31.
    [9] Scott M,McCluggage WG,Hillan KJ,et al.Altered patterns of transcription of the septin gene,SEPT9,in ovarian tumorigenesis[J].Int J Cancer,2006,118(5):1325-9.
    [10] Connolly D,Yang Z,Castaldi M,et al.Septin 9 isoform expression,localization and epigenetic changes during human and mouse breast cancer progression[J].Breast Cancer Res,2011,13(4):R76.
    [11] Tóth K,Galamb O,Spisák S,et al.The influence of methylated septin 9 gene on RNA and protein level in colorectal cancer[J].Pathol Oncol Res,2011,17(3):503-9.
    [12] Amir S,Golan M,Mabjeesh NJ.Targeted knockdown of SEPT9_v1 inhibits tumor growth and angiogenesis of human prostate cancer cells concomitant with disruption of hypoxia-inducible factor -1 pathway[J].Mol Cancer Res, 2010,8(5):643-52.
    [13] Mcllhatton MA,Burrows JF,Donaghy PG,et al.Genomic organization,complex splicing pattern and expression of a human septin gene on chromosome 17q25.3[J].Oncogene,2001,20(41):5930-9.
    [14] Kalikin LM,Sims HL,Petty EM.Genomic and expression analyses of alternatively spliced transcripts of the MLL septin-like fusion gene(MSF) that map to a 17q25 region of loss in breast and ovarian tumors[J].Genomics,2000,63(2):165-72.
    [15] Grützmann R,Molnar B,Pilarsky C,et al.Sensitive detection of colorectal cancer in peripheral blood by septin 9 DNA methylation assay[J].PLoS One,2008,3(11):e3759.
    [16] deVos T,Tetzner R,Model F,et al.Circulating methylated SEPT9 DNA in plasma is a biomarker for colorectal cancer[J].Clin Chem,2009,55(7):1337-46.
    [17] Bennett KL,Karpenko M,Lin MT,et al.Frequently methylated tumor suppressor genes in head and neck squamous cell carcinoma[J].Cancer Res,2008,68(12):4494-9.
    [18] Montagna C,Lyu MS,Hunter K,et al.The Septin 9 (MSF) gene is amplified and overexpressed in mouse mammary gland adenocarcinomas and human breast cancer cell lines[J].Cancer Res,2003,63(9): 2179-87.
    [19] Gonzalez ME,Peterson EA,Privette LM,et al.High SEPT9_v1 expression in human breast cancer cells is associated with oncogenic phenotypes[J].Cancer Res,2007,67(18):8554-64.
    [20] Amir S,Wang R,Matzkin H,et al.MSF-A interacts with hypoxia-inducible factor-1alpha and augments hypoxia-inducible factor transcriptional activation to affect tumorigenicity and angiogenesis[J].Cancer Res,2006,66(2):856-66.
计量
  • 文章访问数:  1472
  • HTML全文浏览量:  20
  • PDF下载量:  369
  • 被引次数: 0
出版历程
  • 收稿日期:  2012-05-27
  • 修回日期:  2012-08-14
  • 刊出日期:  2013-03-24

目录

    /

    返回文章
    返回
    x 关闭 永久关闭