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HCCR-2干扰真核表达载体的构建及其稳定转染细胞系的建立

郭军, 房殿春, 杨仕明, 熊震, 汤旭东, 罗元辉

郭军, 房殿春, 杨仕明, 熊震, 汤旭东, 罗元辉. HCCR-2干扰真核表达载体的构建及其稳定转染细胞系的建立[J]. 实用临床医药杂志, 2008, (1): 27-30. DOI: 10.3969/j.issn.1672-2353.2008.01.005
引用本文: 郭军, 房殿春, 杨仕明, 熊震, 汤旭东, 罗元辉. HCCR-2干扰真核表达载体的构建及其稳定转染细胞系的建立[J]. 实用临床医药杂志, 2008, (1): 27-30. DOI: 10.3969/j.issn.1672-2353.2008.01.005
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GUO Jun, FANG Dian-chun, YANG Shi-ming, XIONG Zhen, TANG Xu-dong, LUO Yuan-hui. Construction of RNAi recombinant eukaryotic expression vectors of HCCR-2 and establishment of its stably transfected cell lines[J]. Journal of Clinical Medicine in Practice, 2008, (1): 27-30. DOI: 10.3969/j.issn.1672-2353.2008.01.005
Citation: GUO Jun, FANG Dian-chun, YANG Shi-ming, XIONG Zhen, TANG Xu-dong, LUO Yuan-hui. Construction of RNAi recombinant eukaryotic expression vectors of HCCR-2 and establishment of its stably transfected cell lines[J]. Journal of Clinical Medicine in Practice, 2008, (1): 27-30. DOI: 10.3969/j.issn.1672-2353.2008.01.005
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HCCR-2干扰真核表达载体的构建及其稳定转染细胞系的建立

  • 第三军医大学西南医院全军消化病研究所,重庆,400038

基金项目: 国家自然科学基金
详细信息

  • 中图分类号: R394.18

Construction of RNAi recombinant eukaryotic expression vectors of HCCR-2 and establishment of its stably transfected cell lines

摘要

    摘要
  • 摘要: 目的 构建HCCR-2于扰真核表达载体,获得干扰质粒稳定转染的HepG2细胞系.方法 人工合成HCCR-2基因干扰序列并定向插入到RNAi真核表达载体PGenesil-l,通过测序鉴定.将干扰质粒用脂质体法转染HeoG2细胞,经G418持续压力选择和有限稀释法获得稳定转染的细胞系.RT-PCR检测筛选克隆HCCR-2 mRNA的表达水平.结果 测序表明HCCR-2干扰序列及读框完全正确,干扰质粒稳定转染的HepG2细胞系在倒置荧光显微镜下呈绿色荧光.RT-PCR结果显示RNAi-H1、RNAi-H3序列对HCCR-2 mRNA有较好的抑制效果.结论 成功构建了HCCR-2干扰真核表达载体,HCCR-2干扰质粒稳定转染的HepG2细胞系的建立为进一步研究HCCR-2在肝癌细胞中的作用奠定了基础.
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  • Ko J, Lee Y H, Hwang S Y. Identification and differential expression of novel human cervical cancer oncogene HOCR-2 in htmam cancers and its involvement in p53 stabilization [J]. Oncogene, 2003, (30):4679.doi: 10.1038/sj.onc.1206624.
    Chung Y J, Kim J W. Novel oneogene HCCR:itsdiagnostic and therapeutic implications for cancer [J]. Histology and Histopathology, 2005(3):999.
    Ko J, Shin S M, Oh Y M, et a1. Transgenic mouse model for breast cancer:induction of breast cancer in novel oncogene HCCR-2 transgenic mice [J]. Oncogene, 2004, (10):1950.doi: 10.1038/sj.onc.1207356.
    Jung S S, Park H S, Lee I J. The HCCR oncoprotein as a biomarker for human breast cancer [J]. Advances in Cancer Research, 2005, (21):7700.
    Yoon S K, Lira N K, Ha S A. The human cervical csncer oncogene protein is a biomarker for hiumn hepatocelMar carcinoma [J]. Cancer Research, 2004, (15):5434.doi: 10.1158/0008-5472.CAN-03-3665.
    杨杨, 张国新, 施瑞华. 人宫颈癌基因蛋白在肝细胞癌中的表达及其临床意义 [J]. 中华肝脏病杂志, 2007(3):223.doi: 10.3760/j.issn:1007-3418.2007.03.018.
    郭军, 房殿春, 姜锋. 人子宫颈癌基因HCCR-2真核表达载体的构建及鉴定 [J]. 第四军医大学学报, 2007(3):250.doi: 10.3321/j.issn:1000-2790.2007.03.017.
    Chakraborty C. Potentiality of small interfering RNAs(siRNA)as recent therapeutic targets for gene-silencing [J]. Current Drug Targets, 2007(3):469.doi: 10.2174/138945007780058988.
    Haney S A. Expanding the repertoire of RNA interference screens for developing new anticancer drug targets [J]. EXPERT OPINION ON THERAPEUTIC TARGETS, 2007, (11):1429.
    Li Y L, Quarles L D, Zhoi H H. RNA interference and its application in bone-rdated diseases [J]. Biochemical and Biophysical Research Communications, 2007(4):817.doi: 10.1016/j.bbrc.2007.07.123.
    de la Fuente J, Kocan K M, Aknazánc. RNA interference for the study and genetic manipulation of ticks [J]. Trends Pmasitol, 2007(9):427.
    Kim D H, Rossi J J. Strategies for silencing hmnan disease using RNA intederence [J]. Nature Reviews Genetics, 2007(3):173.doi: 10.1038/nrg2006.
    Snove O Jr, RossiJ J. Chemical modifications rescue off-target effects of RNAi [J]. ACS Chemical Biology, 2006(5):274.doi: 10.1021/cb6002256.
    Qiu S, Lame T. RNA string kernels for RNAi off-target evaluation [J]. International Journal of Bioinformatics Research and Applications, 2006(2):132.
    Masiero M, Nardo G, Indracoolo S. RNA interference:implications for cancer treatment [J]. Molecular Aspects of Medicine, 2007(1):143.doi: 10.1016/j.mam.2006.12.004.
    Pai S I, Lin YY, Macaes B. Prospects of RNA interference therapy for cancer [J]. Gene Therapy, 2006(6):464.doi: 10.1038/sj.gt.3302694.
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  • 发布日期:  2008-07-13

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