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c-Myc通过缺氧诱导因子-1α调控结肠癌细胞HT-29的增殖

ISSN号：1004-0781
期刊名称：医药导报
时间：2013.4.1
页码：438-442
分类：R735.3[医药卫生—肿瘤;医药卫生—临床医学]
作者机构：[1]华中科技大学同济医学院附属同济医院胃肠外科,武汉430030
相关基金：国家973计划资助项目（2009CB521802）;国家自然科学基金资助项目（30872472,30973496,30800569）
相关项目：miR-363介导DAB2IP调控前列腺癌上皮间质化和肿瘤干细胞抑制肿瘤复发、转移

关键词：结肠癌, C-MYC, 缺氧诱导因子-1α, 细胞增殖, Colon carcinoma, c-Myc , HIF-1 α , Cell proliferation

中文摘要：

目的研究癌基因c-Myc对人结肠癌细胞HT-29中缺氧诱导因子-1α（HIF-1α）的影响及其对肿瘤细胞增殖的作用. 方法构建高表达c-Myc的质粒pcDNA3.1-c-Myc;将构建好的pcDNA3.1-c-Myc质粒转染入HT-29细胞中, 通过Western blot印迹法检测c-Myc在HT-29细胞中的表达情况. 应用Real-time PCR和Western blot印迹法检测常氧和乏氧状态下HT-29细胞系中HIF-1α的mRNA和蛋白水平, 同时检测HIF-1α下游靶基因血管内皮生长因子（VEGF）、内皮型一氧化氮合酶（eNOS）的mRNA水平. 共转染pcDNA3.1-c-Myc质粒和HIF-1α的siRNA入HT-29细胞中. 应用噻唑蓝（MTT）法检测HT-29细胞的增殖情况. 结果成功构建pcDNA3.1-c-Myc质粒;常氧、乏氧状态下转染pcDNA3.1-c-Myc质粒后, HT-29细胞系中HIF-1α的mRNA水平无变化, VEGF和eNOS的mRNA水平明显升高, HIF-1α蛋白水平明显增高. 高表达c-Myc的HT-29细胞增殖能力明显增强. 共转染pcDNA3.1-c-Myc质粒和HIF-1α的siRNA后, HT-29细胞的增殖能力明显降低. 结论在HT-29细胞系中c-Myc通过HIF-1α调控HT-29细胞的增殖.

英文摘要：

Objective To investigate the influence of c-Myc on hypoxia-inducible factor 1, α subunit （ HIF-1 α） and proliferationof HT-29 cells. Methods The plasmid pcDNA3. 1-c-Myc was constructed and transfected into human colon carcinoma cell line HT-29. Efficiency of transfection was detected by Western blotting. The mRNA and protein levels of HIF-1α in normoxia and hypoxia were measured by real-time PCR and Western blotting. The mRNA levels of vascular endothelial growth factor （VEGF） and eNOS （the target genes of HIF-1α） were measured by real-time PCR. PcDNA3.1-c-Myc plasmid and HIF- 1 α siRNA were co-transfeeted into HT-29 ceils. MTF was used to detect the proliferation of the HT-29 cells. Results The plasmid pcDNA3.1-c-Mye was successfully constructed. After transfection into HT-29 cells in normoxia and hypoxia, real-time PCR showed that the mRNA level of HIF-lα had no change, while Western blotting showed the protein level of HIF-lα was significantly increased. The mRNA levels of VEGF and eNOS were significantly increased. The proliferation of HT-29 cells was stimulated through the over-expression of c-Myc. But co-transfection of pcDNA3.1-c-Myc plasmid and HIF-1 α siRNA decreased cell proliferation in HT-29 cell line. Conclusion c-Myc regulates the proliferation of HT-29 cells through HIF-1 α.

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