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编号:10856857
抑癌基因p27对肾癌细胞系GRC┐1生长的抑制作用
http://www.100md.com 《第四军医大学学报》 2001年第17期
肾癌,,肾癌;p27基因;基因表达;基因转染,1材料和方法,2结果,3讨论,参考文献
     关键词: 肾癌;p27基因;基因表达;基因转染

    中图号:Q228

    摘 要:目的 探讨p27基因对肾癌GRC-1细胞系生长的抑制作用,为肾癌发生机制的研究和基因治疗提供理论依据. 方法 采用基因转染技术,将p27cDNA转染到肾癌GRC-1细胞系中,了解其对细胞增殖能力及细胞周期的影响. 结果 转染p27基因的肾癌细胞生长受到抑制,其在软琼脂上克隆形成能力降低了大约4倍.对细胞周期分析表明,处在G0 -G1 期的细胞由32.2%增加到66.9%,经Dot blot和Western blot杂交证实,p27mRNA表达有明显差异(P<0.01),p27的蛋白表达量有明显差异(P<0.01).说明p27蛋白具有生长抑制功能,其作用点为G1 -S. 结论 提高肾癌细胞中p27的表达,能抑制肿瘤细胞的生长,导入p27基因是一种治疗肾癌的新途径.

    p27gene suppresses growth of human renal carcino┐ma GRC┐1cells

    ZHANG Yun-Tao,LIU Fan,JIANG Ru,NAN Peng-Juan,WANG Yong,YU Wen-Qiang,LIU Rong-Fu,LI Yu-Mei

    Department of Urology Surgery,Tangdu Hospital,Fourth Military Medical University,Xi'an710038,China,Department of Chemistry,Faculty of Pre-clinical Medicine,Department of Hematology,Xi-jing Hospital,Fourth Military Medical University

    Keywords:renal neoplasms;p27gene;gene expression;gene transfection

    Abstract:AIM To explore the effect of p27gene on the growth inhibition of renal carcinoma cell line GRC-1.METHODS The p27cDNA was transfected into human re-nal carcinoma cell line GRC-1cells with lipofectamine.The growth rate,cell cycles and colony formation rate of trans-fected cells were observed by means of MTT and FCM assay.RESULTS Expression of p27in GRC-1was identified by Dot blot and Western blot analyses.The growth rate of GRC-1transfected with p27gene was markedly suppressed.Colony formation in soft agar was also decreased significant-ly.Cell cycle analysis by flow cytometry show that the num-ber of cells in G0 -G1 phase of GRC-1cells was significantly increased while cells in S and G2 +M phase was decreased compared to that of the control GRC-1cells.CONCLUSION Transduction of p27gene into lower expression cancer cells can restore its suppressive effect on cell growth by arrest of cell cycle at G1 phase. ......

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