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HLCDG1基因转染对肺癌细胞生长的影响 被引量:3
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作者 邹飞雁 谢海龙 +3 位作者 陈主初 何春梅 关勇军 李友军 《癌症》 SCIE CAS CSCD 北大核心 2003年第11期1121-1126,共6页
背景与目的:HLCDGI基因是我们实验室最近克隆的新基因,它位于5q33,介于D5S436~D5S470之间,在肺癌组织中明显表达下调或缺失。本研究旨在观察HLCDGI基因是否具有抑制肺癌细胞生长的能力。方法:构建HLCDGI基因的重组质粒pcDNA3.1(+)/HLCD... 背景与目的:HLCDGI基因是我们实验室最近克隆的新基因,它位于5q33,介于D5S436~D5S470之间,在肺癌组织中明显表达下调或缺失。本研究旨在观察HLCDGI基因是否具有抑制肺癌细胞生长的能力。方法:构建HLCDGI基因的重组质粒pcDNA3.1(+)/HLCDGI,通过脂质体转染,将HLCDGI基因cDNA导入肺癌细胞系A549中,经G418筛选,获得稳定表达的细胞克隆。RT-PCR检测HLCDGI基因表达,并采用细胞生长抑制实验、软琼脂集落形成实验及裸鼠致瘤性实验,分析HLCDGI基因转染细胞恶性表型。结果:RT-PCR结果表明转染HLCDGI 基因的细胞HLCDGI mRNA明显表达。转染HLCDGI基因组、转染空载体组和未转染组,3组细胞生长倍增时间分别为70.0 h、43.3 h和39.5 h,转染HLCDGI基因组与两对照组之间的差异有显著性(P<0.05)。计算软琼脂中细胞克隆形成率,结果分别为8.5%、29.0%和35.0%,转染HLCDGI基因的细胞克隆形成率明显降低。将转染HLCDGI基因的细胞、转染空载体的细胞和未转染的细胞分别注射入无胸腺的裸鼠体内,43天后处死动物,剥离出肿瘤组织并称重,各组瘤块平均重量分别为0.120g、0.612g和0.924g,转染HLCDGI基因的细胞成瘤能力与两对照组细胞比较,差异有显著性(P<0.05)。结论:HLCDGI在肺癌A549细胞中表达。 展开更多
关键词 hlcdg1基因 基因转染 肺癌 细胞生长 检测 基因表达
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肺癌候选抑瘤基因HLCDG1片段的验证及全长序列的克隆 被引量:2
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作者 邹飞雁 谢海龙 +3 位作者 余艳辉 欧阳咏梅 贺智敏 陈主初 《中国病理生理杂志》 CAS CSCD 北大核心 2009年第6期1081-1088,共8页
目的:对肺癌候选抑瘤基因HLCDG1片段(已知序列)进行验证,并克隆其全长cDNA序列。方法:采用RT-PCR和末端快速扩增技术(RACE),对正常人肺组织中HLCDG1基因片段进行验证和克隆其全长cDNA序列,登录基因数据库进行比对分析。结果:从正常人肺... 目的:对肺癌候选抑瘤基因HLCDG1片段(已知序列)进行验证,并克隆其全长cDNA序列。方法:采用RT-PCR和末端快速扩增技术(RACE),对正常人肺组织中HLCDG1基因片段进行验证和克隆其全长cDNA序列,登录基因数据库进行比对分析。结果:从正常人肺组织中钓取到HLCDG1基因未知的3’端和5’端序列分别为1 304 bp和1 548 bp。HLCDG1基因与人酪蛋白激酶Ⅰα(CSNK1A1)基因2号转录变体相似性达99%,score=5 544,E=0。结论:HLCDG1基因为已知的CSNK1A1基因的2号转录变体,可能参与肺癌发生发展。 展开更多
关键词 基因 hlcdg1 基因 CSNK1A1 DNA末端快速扩增法 肺肿瘤
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HLCDG1基因siRNA表达质粒的构建及其对A549细胞周期和增殖的影响 被引量:1
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作者 邹飞雁 李峰 +4 位作者 陈主初 贺智敏 吕辉 刘孝荣 欧阳咏梅 《中国病理生理杂志》 CAS CSCD 北大核心 2006年第7期1256-1262,共7页
目的:本研究旨在应用RNA干扰技术,诱导转基因肺癌细胞HLCDG1基因沉默并观察其对肺癌细胞周期和增殖的影响。方法:构建HLCDG1基因短小干扰双链RNA表达载体,筛选HLCDG1基因表达沉默的肺癌细胞株并应用MTT和流式细胞仪分析这一肺癌细胞株... 目的:本研究旨在应用RNA干扰技术,诱导转基因肺癌细胞HLCDG1基因沉默并观察其对肺癌细胞周期和增殖的影响。方法:构建HLCDG1基因短小干扰双链RNA表达载体,筛选HLCDG1基因表达沉默的肺癌细胞株并应用MTT和流式细胞仪分析这一肺癌细胞株增殖和细胞周期分布状况。结果:根据siRNA表达载体的设计原则,构建了能在哺乳动物细胞稳定表达的siRNA载体5个(4个位点匹配实验组和1个位点错配对照组),依次命名为pHL-si-1、pHL-si-2、pHL-si-3、pHL-si-4和pHL-si-c。上述载体分别与表达HLCDG1的重组体pcDNA3.1(+)/HLCDG1共转染A549细胞,筛选到一株共转染pHL-si-1载体和pcDNA3.1(+)/HLCDG1重组体的A549细胞,它几无HLCDG1基因表达,这株细胞暂命名为A549-HLCDG1-si-1。MTT和流式细胞仪分析表明,A549-HLCDG1-si-1细胞增殖能力明显提高,进入S期和G2+M期增多,滞留G1期减少。结论:采用RNAi技术特异阻断HLCDG1基因表达,验证了HLCDG1基因对A549肺癌细胞有生长抑制作用。 展开更多
关键词 基因 hlcdg1 短小干扰双链RNA A549细胞 细胞周期
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Identification and significance of differential proteins in A549 cells transfected with HLCDG1
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作者 ZOU Fei-yan XIE Hai-long ZENG Ping-yao CHEN Zhu-chu LI Feng XIAO Zhi-qiang FENG Xue-ping ZHANG Peng-fei YANG Hai-yan HU Wei YU Yan-hui OUYANG Yong-mei 《Journal of Central South University of Technology》 2005年第z1期337-342,共6页
HLCDG1, which locates in chromosome 5q33, is a novel gene cloned recently. The HLCDG1 expression was significantly down regulated in the primary lung carcinoma. It was previously studied that HLCDG1 acted like a tumor... HLCDG1, which locates in chromosome 5q33, is a novel gene cloned recently. The HLCDG1 expression was significantly down regulated in the primary lung carcinoma. It was previously studied that HLCDG1 acted like a tumor suppressor gene. In this paper, proteomics studies were performed to analyze the proteomic expression patterns in the HLCDG1-transfected human lung carcinoma cell line (A549-HLCDG1) and in the control vector-transfecred human lung carcinoma cell line (A549-vector). Employing two dimensional gel eleetrophoresis (2DE), the global pattern of protein expressions in A549-HLCDG1 human lung adenocarcinoma cell line expressing stably HL-CDG1 gene were compared with those of control A549-vector cell line to generate a differential protein expression catalog. Forty-two differentially expressed proteins were screened. Thirteen differential proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), which were 6 upregulated (MSH5, MOD, MDH precursor, ETFβ, Prxd Ⅵ and JM23) and 7 downregulated (PLC-δ1, hnRNPA2,hnRNPB1, TIM, TCTP, nm23H-1 and PrxdⅤ) proteins in A549-HLCDG1 cells compared to control A549-vector cells. The above identified proteins were involved in energy metabolism, transcription regulation, antioxidation,cell cycle, metastasis, DNA methylation and mismatch repair. Therefore, these differential expression proteins by HLCDG1 transfection may play some important roles for investigation of the biochemical basis of growth suppression of HLCDG1 gene in lung carcinoma cells A549. Further understanding of this data base may provide valuable resources for the developing novel diagnostic markers and therapeutic targets of lung cancer. 展开更多
关键词 hlcdg1 gene LUNG carcinoma cell two-dimensional GEL ELECTROPHORESIS mass SPECTROMETRY PROTEOMICS
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