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锌带基因ZNRD1在胃癌耐药细胞中的表达和功能 被引量：6

Expression and function of zinc ribbon gene ZNRD1 in drug-resistant gastric cancer cells
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摘要 目的　探讨锌带基因ZNRD1在多药耐药胃癌细胞中的表达和作用。方法　应用Northernblot和半定量RT PCR ,观察锌带基因ZNRD1在胃癌细胞SGC790 1及其长春新碱 (VCR)诱导的耐药细胞SGC790 1/VCR中的表达 ;将反义核酸转染SGC790 1/VCR细胞 ,通过免疫组化检测转导细胞与非转导细胞ZNRD1蛋白的表达 ;以流式细胞仪检测细胞周期的变化 ,以MTT实验检测细胞生长曲线和对VCR、阿霉素 (ADM)的药敏性。结果　胃癌耐药细胞SGC790 1/VCR与非耐药细胞相比 ,ZNRD1基因的表达明显增高。转导株antiZNRD1 SGC790 1/VCR细胞的ZNRD1蛋白水平明显低于非转导株 ,G1期细胞比例增加而S期比例减少 ,生长受到抑制 ,且对VCR、ADM的敏感性明显增高。结论　胃癌耐药细胞与非耐药细胞相比 ,ZNRD1基因处于高表达状态。反义核酸转染可有效阻断ZNRD1蛋白的表达 ,在一定程度上逆转人胃癌耐药细胞SGC790 1/VCR的多药耐药性。 Objective To study the expression and function of zinc ribbon gene ZNRD1 in drug resistant cells of gastric cancer. Methods Two tumor cell lines were used in this study: gastric cancer SGC7901 and its drug resistant counterpart SGC7901/VCR stepwise selected by vincristine. The expression of ZNRD1 in SGC7901 and SGC7901/VCR was detected by northern blot and semiquantitative RT PCR. ZNRD1 antisense nucleic acid was transfected into SGC7901/VCR cells by lipofectamine. The expression of protein in SGC7901/VCR cells and the transfectants was detected by immunochemical method. Fluorescence activated cell scan (FACS) was applied to observe the cell cycle alteration. Growth curve and drug sensitization of cells for vincristine (VCR) and adriamycin (ADM) were analyzed by MTT assay. Results The expression of ZNRD1 was higher in SGC7901/VCR than in SGC7901 cells. Immunochemical results showed that the expression level of ZNRD1 protein was lower in anti ZNRD1 SGC7901/VCR cells than in non transfectants. The anti ZNRD1 SGC7901/VCR cells were gradually accumulated in G 1 phase, with a concomitant decrease of cell population in S phase. MTT assay showed that transfectant cell proliferation was lagged and more sensitive to VCR and ADM than non transfectants. Conclusion ZNRD1 gene displays high expression in VCR resistant gastric cancer cells. Expression of ZNRD1 protein is effectively blocked in anti ZNRD1 SGC7901/VCR cells by gene transfection. ZNRD1 antisense nucleic acid could reverse, to some degree, the MDR of human drug resistant gastric cancer cell SGC7901/VCR.
作者 张宇梅 赵燕秋 严泉剑 潘阳 林易辉 樊代明
机构地区 第四军医大学西京医院消化病研究所
出处 《中华肿瘤杂志》 CAS CSCD 北大核心 2003年第2期125-129,共5页 Chinese Journal of Oncology
基金 国家自然科学基金重点资助项目 ( 3 0 0 3 0 14 0 ) 创新研究群体基金资助项目 ( 3 0 0 2 40 0 2 )
关键词 胃癌 多药耐药性 锌带基因ZNRD1 反义核酸 SGC7901细胞 Stomach neoplasms Zinc ribbon gene Antisense nucleic acid Multidrug resistance
分类号 R735.2 [医药卫生—肿瘤]
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参考文献9

  • 1张宇梅,赵燕秋,金晓航,时永全,乔泰东,陈宝军,樊代明.ZNRD1基因的克隆及其反义核酸对胃癌耐药细胞阿霉素蓄积的影响[J].解放军医学杂志,2002,27(4):330-332. 被引量：7
  • 2Chen HT, Legault P, Glushka J, et al. Structure of a (Cys3His) zinc ribbon, a ubiquitous motif in archaeal and eucaryal transcription. Protein Sci, 2000, 9: 1743-1752.
  • 3Hem ming SA, Jansma DB, Macgregor PF, et al. RNA polymerase Ⅱ subunit Rpb9 regulates transcription elongation in vivo. J Biol Chem,2000, 275: 35506-35511.
  • 4McCoy C, McGee SB, Cornwell MM. The Wilms' tumor suppressor,WT1, inhibits 12-O-tetradecanoylphorbol-13-acetate activation of the multidrug resistance-1 promoter. Cell Growth Differ, 1999, 10: 377-386.
  • 5Hirose M. The role of Wilms' tumor genes. J Med Invest, 1999, 46:130-140.
  • 6Bartsevich VV, Juliano RL. Regulation of the MDR1 gene by transcriptional repressors selected using peptide combinatorial libraries.Mol Pharmacol, 2000 , 58: 1-10.
  • 7Frick DN, Richardson CC. DNA PRIMASES. Biochem, 2001,70:39-80.
  • 8Zhao Y, You H, Liu F, et al. Differentially expressed gene profiles between multidrug resistant gastric adenocarcinoma cells and their parental cells. Cancer Lett, 2002, 185: 211.
  • 9Fan W, Wang Z, Kyzysztof F, et al. A new zinc ribbon gene (ZNRD1)is cloned from the human MHC class Ⅰ region. Genomics, 2000, 63:139-141.

二级参考文献9

  • 1[1]Fan W,Wang Z,Kyzysztof F et al.A new zinc ribbon gene (ZNRD1) is cloned from the human MHC class I region.Genomics,2000,63(1):139
  • 2[2]Fan W,Cai W,Parimoo S et al.Identification of seven new human MHC class I region genes around the HLA-F locus.Immunogenetics,1996,44(2):97
  • 3[3]Qian X,Jeon C,Yoon H et al.Structure of a new nucleic-acid-binding motif in eukaryotic transcriptional elongation factor TFIIS.Nature,1993,365(6443):277
  • 4[4]Chu TW,Capossela A,Coleman R et al.Cloning of a new "finger" protein gene (ZNF173) within the class I region of the human MHC.Genomics,1995,29(1):229
  • 5[5]Chen HT,Legault P,Glushka J et al.Structure of a (Cys3His) zinc ribbon,a ubiquitous motif in archaeal and eucaryal transcription.Protein Sci,2000,9(9):1743
  • 6[6]Grishin NV.C-terminal domains of Escherichia coli topoisomerase I belong to the zinc-ribbon superfamily.J Mol Biol,2000,299(5):1165
  • 7[7]McCoy C,McGee SB,Cornwell MM.The Wilms' tumor suppressor,WT1,inhibits 12-O-tetradecanoylphorbol-13-acetate activation of the multidrug resistance-1 promoter.Cell Growth Differ,1999,10(6):377
  • 8[8]Hirose M.The role of Wilms' tumor genes.J Med Invest,1999 ,46(3-4):130
  • 9[9]Bartsevich VV,Juliano RL.Regulation of the MDR1 gene by transcriptional repressors selected using peptide combinatorial libraries.Mol Pharmacol,2000,58(1):1

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中华肿瘤杂志
2003年 第2期

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