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PinX1基因真核表达载体的构建及其对乳腺癌MCF-7细胞增殖的抑制作用

Construction of Recombinant Eukaryotic Expression Vector for PinX1 and Its Roles in Inhibiting Proliferation of Human Breast Cancer MCF-7 Cell Line
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摘要 探讨了PinX1基因在乳腺癌MCF-7细胞生长和细胞周期中的作用,初步探讨了该基因用于乳腺癌临床治疗的可行性.采用RT-PCR技术从293-T细胞中扩增PinX 1基因,将其克隆入真核表达载体pEGFP-C1中,再将重组质粒转染MCF-7细胞.通过real-time PCR检测PinX1基因的mRNA表达,用MTT法检测转染前后细胞生长曲线的变化,用流式细胞仪检测转染目的基因后细胞生长周期的改变.检测结果表明,PinX1基因已经在转染后MCF-7细胞的细胞核内稳定表达,乳腺癌细胞生长明显减缓(P<0.05),增殖变慢(P<0.05),细胞生长阻滞于G0/G1期,说明PinX1基因可抑制乳腺癌MCF-7细胞的生长和增殖. This paper explores the effects of PinX1 gene on the growth and the cell cycles of breast carcinoma cell line MCF-7 and the potential in treatment of breast cancer. The PinX1 mRNA was amplified from 293-T cell with RT-PCR and inserted into pEGFP-C1 vector. The recombined eukaryotic expression vector of PinX1 was confirmed by enzyme digestion, and transfected into the breast carcinoma cell line MCF-7 using a liposome method. The expressed mRNA was detected by real-time PCR, and the cell growth and cell cycles by MTT and flow cytometry, respectively. Results showed that PinX1 transfection into the breast cancer MCF-7 cell line was stable and the transfected gene was integrated into nucleolus DNA of transfected cells. Transfection of PinX1 gene could significantly inhibit the growth and proliferation of breast cancer cells (P 〈 0.05) and the cell growth was arrested at G0/G1 stage. The conclusion is that PinX1 gene could inhibit growth and proliferation of breast cancer cells.
出处 《上海大学学报(自然科学版)》 CAS CSCD 北大核心 2013年第6期631-635,共5页 Journal of Shanghai University:Natural Science Edition
基金 国家自然科学基金资助项目(10972130)
关键词 乳腺癌 PinX1基因 载体构建 细胞增殖 breast cancer PinX1 gene vector construction cell proliferation
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参考文献16

  • 1BLASCO M A. Telomeres and human disease:ageing,cancer and beyond[J].{H}Nature Reviews Genetics,2005,(08):611-622.
  • 2ZHOU X Z,LU K P. The Pin2/TRF1 interacting protein PinX1 is a potent telomerase inhibitor[J].{H}CELL,2001,(03):347-359.
  • 3LIAO C,ZHAO M J,SONG H. Identification of the gene for a novel liver-related putative tumor suppressor at a high frequency loss of heterozygosity region of chromosome 8p23 in human hepatocellular carcinoma[J].{H}HEPATOLOGY,2000.721-722.
  • 4孙文旦.PinX1基因真核表达载体的构建及其在Hek293细胞中的表达[J].临床检验杂志,2009,27(6):445-447. 被引量:1
  • 5WON S P,JONG H L,JIK Y P. Genetic analysis of the liver putative tumor suppressor gene in hepatocelluar carcinomas[J].{H}Cancer letters,2002,(02):199-207.
  • 6DE LANGE T. Shelterin:the protein complex that shapes and safeguards human telomeres[J].{H}Genes and development,2005,(18):2100-2110.
  • 7VAN STEENSEL B,DE LANGE T. Control of telomere length by the human telomeric protein TRF1[J].{H}NATURE,1997.740-743.
  • 8CHONG L,VAN S B,BROCCOLI D. A human telomeric protein[J].{H}SCIENCE,1995,(5242):1663-1667.doi:10.1126/science.270.5242.1663.
  • 9ANCELIN K,BRUNORI M,BAUWENS S. Targeting assay to study the cis functions of human telomeric proteins:evidence for inhibition of telomerase by TRF1 and for activation of telomere degradation by TRF2[J].{H}Molecular and Cellular Biology,2002,(10):3474-3487.
  • 10SOOHOO C Y,SHI R,LEE T H. Telomerase inhibitor PinX1 provides a link between TRF1 and telomerase to prevent telomere elongation[J].{H}Journal of Biological Chemistry,2011.3894-3906.

二级参考文献8

  • 1Zhou X Z, Lu K P. The Pin2/TRF1-interacting protine PinX1 is a potent telomerase inbibtor[ J]. Cell ,2001,107:347-359.
  • 2Won S P,Jong H L,Jik Y P,et al. Genetic analysis of the liver putative tumor suppressor gene in hepatocellular carcinomas [ J ]. Cancer Let,2002,175 (2) : 199-207.
  • 3Liao C, Zhao M J, Song H, et al. Identification of the gene for a novel liver-related putative tumor suppressor at a high frequeny loss of heterozygosity region of chromosome 8p23 in human hepatocellular careinoma [ J ]. Hepatology,2000,10:721-722.
  • 4Frederick M. Short Protocols in Molecular Biology [M]. 3rd ed. John Wiley and Sons,Inc. ,1995:277-278.
  • 5Chang Q, Pang J C, Li J, et al. Molecular analysis of PinX1 in medulloblastomas [ J ]. Int J Cancer,2004,109 (2) :309-314.
  • 6Akiyama Y, Maesawa C, Wada K, et al. Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma[ J]. Oncol Rep,2004,11 (4) :871-874.
  • 7Yoo J E, Oh B K, Park Y N,et al. PinX1 mediates TRF1 accumulation in nucleolus and enhances TRF1 binding to telomeres [J]. J Mol Biol, 2009,388 ( 5 ) :928-940.
  • 8Oh B K,Yoon S M,Lee C H,et al. Rat homolog of PinX1 is a nucleolar protein involved in the regulation of telomere length [J]. Gene, 2007,400(1-2) :35-43.

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