| 重组人源化ING4基因的构建及其对肺癌NCI-H460细胞生长抑制作用 |
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| 引用本文： | 段光军,韩志强,林云霞,杨鹏,胡华成,盛伟华,王金志,杨吉成.重组人源化ING4基因的构建及其对肺癌NCI-H460细胞生长抑制作用[J].肿瘤,2008,28(11). |
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| 作者姓名： | 段光军 韩志强 林云霞 杨鹏 胡华成 盛伟华 王金志 杨吉成 |
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| 作者单位： | 1. 苏州大学附属第二医院呼吸内科,苏州,215004
2. 苏州大学基础医学院细胞与分子生物学教研室.苏州,215123 |
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| 摘 要： | 目的:构建重组人源化hING4基因并探究其对肺癌细胞的生长抑制效应.方法:以pcDNA3.0-mING4重组质粒为模板.通过定点突变技术构建带有绿色荧光蛋白(GFP)的重组腺病毒Ad-hING4基因.用荧光显微镜和RT-PCR法检测hING4在肺癌细胞中的表达;Westem印迹法检测hING4对肺癌细胞中Bax、Bcl-2表达的影响;集落形成试验和FCM法检测hING4对肺癌细胞增殖和凋亡影响.结果:基因测序和PCR结果提示成功构建人源化Ad-hING4基因;并能在肺癌细胞中表达,对肺癌细胞有细胞毒作用;Westem印迹法检测结果提示基因在上调Bax表达同时下调Bel-2的表达;集落形成试验、FCM检测结果提示hING4基因可抑制肺癌细胞增殖并诱导凋亡.结论:成功构建了Ad-hING4基因,并能在体外抑制肺癌细胞生长.
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| 关 键 词： | 肺肿瘤 INC4蛋白 人 诱变 突点 基因 肿瘤抑制 |
Construction of recombinant hING4 gene and its inhibitory effect on the growth of lung cancer NCI-H460 cells |
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| DUAN Guang-jun,HAN Zhi-qiang,LIN Yun-xia,YANG Peng,HU Hua-cheng,SHEN Wei-hua,WANG Jin-zhi,YANG Ji-cheng.Construction of recombinant hING4 gene and its inhibitory effect on the growth of lung cancer NCI-H460 cells[J].Tumor,2008,28(11). |
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| Authors: | DUAN Guang-jun HAN Zhi-qiang LIN Yun-xia YANG Peng HU Hua-cheng SHEN Wei-hua WANG Jin-zhi YANG Ji-cheng |
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| Institution: | DUAN Guang-jun1,HAN Zhi-qiang1,LIN Yun-xia1,YANG-Peng1,HU Hua-cheng1,SHEN Wei-hua2,WANG Jin-zhi2,YANG Ji-cheng2(1.Department of Respiration Medicine,Second Affiliated Hospital of Soochow University,Suzhou 215004,China,2.Cell , Molecular Biology Institute,College of Medicine,Soochow University,Suzhou 215123,China) |
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| Abstract: | Objective:To construct a recombinant human inhibitor of growth 4(hING4) gene and observe its growth inhibition effect on lung adenocarcinoma NCI H460 cells.Methods: The GFP-labeled recombinant adenovirus vector hING4(Ad-hING4) was constructed using pcDNA3.0-mING4 plasmid as a template and site-specific mutagenesis technique.RT-PCR and fluorescence microscope were used to detect the expression of hING4 in NCI H460 cells.Bax and Bcl-2 protein expression were detected by Western blotting.Colony formation assay... |
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| Keywords: | Lung neoplasms ING4 protein human Mutagenesis site-directed Genes tumor suppressor |
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