|本期目录/Table of Contents|

[1]伦淑敏.肌细胞增强因子2A基因真核表达质粒的构建及对乳腺癌细胞MCF-7增殖能力的影响[J].天津医科大学学报,2014,20(06):429-432.
 LUN Shu-min.Construction of myocyte enhancer factor 2A eukaryotic expression plasmid and effects on cell proliferation in breast cancer cell line MCF7[J].Journal of Tianjin Medical University,2014,20(06):429-432.
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肌细胞增强因子2A基因真核表达质粒的构建及对乳腺癌细胞MCF-7增殖能力的影响(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
20
期数:
2014年06期
页码:
429-432
栏目:
基础医学
出版日期:
2014-11-20

文章信息/Info

Title:
Construction of myocyte enhancer factor 2A eukaryotic expression plasmid and effects on cell proliferation in breast cancer cell line MCF7
文章编号:
1006-8147(2014)06-0429-04
作者:
伦淑敏
(天津医科大学肿瘤医院肿瘤研究所,国家肿瘤临床医学研究中心,天津市肿瘤防治重点实验室 ,天津 300060)
Author(s):
LUN Shu-min
(Tianjin Medical University Cancer Institute and HospitalNational Clinical Research Center of Cancer, Ministry of Education Key Laboratory of Cancer Prevention and TherapyKey Laboratory of Cancer Prevention and TherapyTianjin 300060 China)
关键词:
肌细胞增强因子2A乳腺癌质粒增殖
Keywords:
myocyte enhancer factor 2Abreast cancerplasmidproliferation
分类号:
R737.9
DOI:
-
文献标志码:
A
摘要:
目的:构建MEF2A基因真核表达重组质粒,并转染乳腺癌细胞株,观察其对细胞增殖能力的影响。方法:实时定量聚合酶链反应(RT-QPCR)检测MCF-7、BT474及MDA-MB-231等3种乳腺癌细胞中MEF2A mRNA的表达。采用全基因合成法合成MEF2A编码区序列,克隆入pcDNA3.1(+)-HA载体,构建重组质粒pcDNA3.1-MEF2A-HA。重组质粒转染乳腺癌细胞,采用Western blot及RT-QPCR检测MEF2A的表达效率。细胞计数实验检测细胞的增殖能力。结果:3株乳腺癌细胞中,MDA-MB-231细胞中MEF2A mRNA表达水平较低。其基因测序结果显示与预期片段大小一致,表明MEF2A重组质粒构建成功。将MEF2A重组质粒瞬时转染MDA-MB-231细胞后, MEF2A表达质粒组与对照组比较,其MEF2A表达升高,细胞增殖能力增强。结论:成功构建MEF2A过表达重组质粒,在MDA-MB-231中过表达MEF2A促进细胞的增殖能力。
Abstract:
Objective: To construct eukaryotic expression plasmid of pcDNA3.1-MEF2A and examine its effect on the proliferation of breast cancer cell line. Methods: The MEF2A mRNA levels were detected in MCF7, BT474 and MDA-MB-231 cells, respectively by real-time quantitative PCR. The full-length coding sequence of MEF2A was amplified by standard RT-QPCR. The amplified。MEF2A was cloned into pcDA3.1 vector. The eukaryotic expression plasmid pcDNA3.1-MEF2A-HA was transfected into breast cancer cells. RT-QPCR and western blot were performed to measure the expression level of MEF2A. Cell counting was performed to evaluate the effects of MEF2A expression on cell proliferation. Results: The MEF2A mRNA expression level of MDA-MB-231 was low in MDA-MB-231 cells. Western blot and RT-QPCR indicated that transfection of MEF2A expression vector into MDA-MB-231 cells resulted in increased mRNA/protein levels of MEF2A. The forced MEF2A expression significantly enhanced cell proliferation of MDA-MB-231. Conclusion: The eukaryotic expression plasmid of pcDNA3.1-MEF2A-HA is constructed successfully. Over-expression of MEF2A enhances proliferation ability of MDA-MB-231cells.



 

参考文献/References:

[1]Parkin D M, Bray F, Ferlay J, et al. Global Cancer statistics, 2002[J]. CA Cancer J Clin, 2005,55(2):74
[2]Black B L, Molkentin J D, Olson E N. Multiple rol es for the MyoD basic region in t ransmission of transcript ional activat ion signals and int eract ion with MEF2[Z].1998:69
[3]Nguyen H T, Bodmer R, Abmayr S M.D-mef2:a drosophila mesoderm-specific MADS boxcontaining gene with a biphasic expression profile[Z].
[4]Mckinsey T, Zhang C L, Olson E N.MEF2:a calcium-dependent regulator of cell division,differentiation and death[Z].2002:40
[5]Seok H Y, Tatsuguchi M, Callis T E, et al. miR-155 inhibits expression of the MEF2A protein to repress skeletal muscle differentiation[J]. J Biol Chem, 2011,286(41):35339
[6]Al Madhoun A S, Mehta V, Li G, et al. Skeletal myosin light chain kinase regulates skeletal myogenesis by phosphorylation of MEF2C[J]. EMBO J, 2011,30(12):2477
[7]Tessier S N, Storey K B. Myocyte enhancer factor-2 and cardiac muscle gene expression during hibernation in thirteen-lined ground squirrels[J]. Gene, 2012,501(1):8
[8]Zhao W. (1),Zhao SP,Peng DQ[J]. The effects of myocyte enhancer factor 2A gene on the proliferation,migration and phenotype of vascular smooth muscle cells.Cell Biochem Funct, 2012,30(2):108
[9]Bai X, Wu L, Liang T, et al. Overexpression of myocyte enhancer factor 2 and histone hyperacetylation in hepatocellular carcinoma[J]. J Cancer Res Clin Oncol, 2008,134(1):83-91
[10]Ishikawa F, Miyoshi H, Nose K, et al. Transcriptional induction of MMP-10 by TGF-beta, mediated by activation of MEF2A and downregulation of class IIa HDACs[J]. Oncogene, 2010,29(6):909
[11]Feng H T, Cheng T, Steer J H, et al. Myocyte enhancer factor 2 and microphthalmia-associated transcription factor cooperate with NFATc1 to transactivate the V-ATPase d2 promoter during RANKL-induced osteoclastogenesis[J]. J Biol Chem, 2009,284(21):14667
[12]Daems C, Martin L A.MEF2 IS RESTRICTED TO THE MALE GONAD AND REGULATES EXPRESSION OF THE ORPHAN NUCLEAR RECEPTOR NR4A1[Z].2014:me20131407

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备注/Memo

备注/Memo:
作者简介 伦淑敏(1988-),女,硕士在读,研究方向:乳腺癌相关基因的转录调控机制;E-mail:lunshuminvs@163.com。
更新日期/Last Update: 2014-11-20