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高表达蛋白hSav1对人胚肾HEK293增殖能力的影响
引用本文:李兆明,董硕,刘韦成,蔡少鑫,罗学来,李小兰,陶德定,严群,龚建平,胡俊波.高表达蛋白hSav1对人胚肾HEK293增殖能力的影响[J].中华实验外科杂志,2010,27(6).
作者姓名:李兆明  董硕  刘韦成  蔡少鑫  罗学来  李小兰  陶德定  严群  龚建平  胡俊波
作者单位:华中科技大学同济医学院附属同济医院胃肠外科/分子医学中心,武汉,430030
基金项目:国家重点基础研究发展规划(973计划),国家自然科学基金,湖北省自然科学基金 
摘    要:目的 观察高表达人Salvadorl(hSav1)蛋白对人胚肾293(HEK293)细胞增殖的影响.方法 构建含有绿色荧光蛋白(GFP)的hSav1质粒GFP-N1-hSav1;将构建好的GFP-N1-hSav1质粒在脂质体Lipofectamine 2000的介导下转染HEK293细胞,荧光显微镜下观察质粒转染效率;分别在转染后12、24、36、48 h通过噻唑蓝(MTT)比色法计算细胞增殖抑制率,抑制率=(对照-空白)-(给药-空白)]/(对照-空白)×100%;转染后36 h加入5-溴-2'-脱氧尿苷(BrdU),通过其掺入比率来显示hSav1对细胞增殖的影响.结果 质粒GFP-N1-hSav1构建成功,转染效率为70%-80%左右,蛋白hSav1明显高表达;MTT结果显示转染质粒后12、24、36、48 h,高表达蛋白hSav1组的细胞增殖抑制率分别为2%、5%、15%、23%,而阴性对照分别为2%、3%、2%、2%;BrdU结果显示,在HEK293细胞中高表达蛋白hSav1后细胞增殖抑制率为(27.3±3.8)%,阴性对照组的细胞增殖抑制率为(12.9±5.3)%,差异有统计学意义(P<0.05).结论 在人胚肾HEK293中高表达蛋白hSav1可明显抑制细胞增殖能力.

关 键 词:细胞增殖  人胚肾细胞

Effects of hSav1 on cell proliferation of human embryonic kidney cell line HEK293
LI Zhao-ming,DONG Shuo,LIU Wei-cheng,CAI Shao-xin,LUO Xue-lai,LI Xiao-Ian,TAO De-ding,YAN Qun,GONG Jian-ping,HU Jun-bo.Effects of hSav1 on cell proliferation of human embryonic kidney cell line HEK293[J].Chinese Journal of Experimental Surgery,2010,27(6).
Authors:LI Zhao-ming  DONG Shuo  LIU Wei-cheng  CAI Shao-xin  LUO Xue-lai  LI Xiao-Ian  TAO De-ding  YAN Qun  GONG Jian-ping  HU Jun-bo
Abstract:Objective To elucidate the effects of human Salvador 1 (hSav1 ) on cell proliferation of human embryonic kidney cell line HEK293. Methods The plasmid CFP-N1-hSav1 was constructed and transfected into HEK293 cells with lipofectamine 2000. The transfection efficiency was detected by fluorescent microscopy. The effects of hSav1 on cell proliferation were measured by MTT and BrdU incorporation. Results The transfection efficiency was about 70% -80%. The MTT results showed that the inhibition rate of cell proliferation in transfected group at the 12th, 24th, 36th, and 48th h was 2% , 5% , 15% , and 23% , respectively;while that in the control group was 2% , 3% , 2% , and 2% respectively. The BrdU incorporation revealed that the BrdU incorporation rate in transfected group (12. 9 ±5. 3)% was significantly lower than in control group (27.3±3.8)% (P<0.05). Conclusion hSav1 is a newly identified protein that can cause cell proliferation inhibition in HEK293 cells.
Keywords:hSav1
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