[1]苏新良,任国胜,王小毅,等.人乳腺癌细胞HYAL1基因真核表达载体的构建及其在MCF-7和ZR-75-30细胞中的表达[J].第三军医大学学报,2008,30(02):146-149.
 SU Xin-liang,REN Guo-sheng,WANG Xiao-yi,et al.Construction of eukaryotic expression vector of human HYAL1 gene and its expression in MCF-7 and ZR-75-30 cells[J].J Third Mil Med Univ,2008,30(02):146-149.
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人乳腺癌细胞HYAL1基因真核表达载体的构建及其在MCF-7和ZR-75-30细胞中的表达(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第02期
页码:
146-149
栏目:
论著
出版日期:
2008-01-30

文章信息/Info

Title:
Construction of eukaryotic expression vector of human HYAL1 gene and its expression in MCF-7 and ZR-75-30 cells
作者:
苏新良任国胜王小毅谭金祥
重庆医科大学附属第一医院内分泌外科
Author(s):
SU Xin-liang REN Guo-sheng WANG Xiao-yi TAN Jin-xiang
Department of General Surgery, The First Affiliated Hospital of Chongqing Medical University
关键词:
透明质酸酶 HYAL1真核表达乳腺癌细胞侵袭
Keywords:
hyaluronidase HYAL1 eukaryotic expression breast cancer cell invasion
分类号:
R394-33;R394.2;R737.9
文献标志码:
A
摘要:
目的 构建人乳腺癌HYAL1基因的真核表达载体并稳定转染HYAL1基因低表达的乳腺癌MCF-7和ZR-75-30细胞株,为进一步探讨HYAL1基因在乳腺癌的侵袭和转移中的作用奠定基础。  方法 采用RT-PCR技术从高表达HYAL1的人乳腺癌细胞株MDA-MB-435S中扩增透明质酸酶HYLAL1基因,并将其插入真核表达载体pcDNA3.1/V5-His-TOPO中,构建的重组表达质粒经脂质体介导转染HYAL1基因低表达的乳腺癌MCF-7和ZR-75-30细胞。  结果 用RTPCR成功地扩增出1条1 332 bp的DNA片段,经限制性内切酶酶切分析和DNA测序证明已经成功构建了人乳腺癌HYAL1基因的真核表达载体。RT-PCR证明转染了重组质粒的乳腺癌MCF-7和ZR-75-30细胞可以高效稳定的表达HYAL1基因,且其穿透细胞外基质的能力增强。  结论 成功构建了人乳腺癌HYAL1基因的真核表达载体,获得了稳定表达人HYAL1基因的MCF-7和ZR-75-30细胞克隆,且其侵袭能力增强。
Abstract:
Objective To construct the eukaryotic expression vector of human HYAL1 gene and obtain MCF-7 and ZR-75-30 cell clones expressing HYAL1 gene stably.   Methods The cDNA encoding HYAL1 gene of human breast cancer was amplified by RT-PCR from the total RNA isolated from human MDA-MB-435S cells and inserted into pcDNA3.1/V5-His-TOPO vector. The recombinant plasmid was transferred into MCF-7 and ZR-75-30 cells.    Results A 1332-bp DNA fragment was successfully amplified from human MDA-MB-435S cell. Restriction enzyme digestion analysis and DNA sequencing showed that HYAL1 gene was inserted into recombinant vector. RT-PCR analysis revealed that HYAL1 gene could be expressed stably in the transfected MCF-7 and ZR-75-30 and it had strong invasive potential.    Conclusion The eukaryotic expression vector of human HYAL1 gene was successfully constructed. MCF-7 and ZR-75-30 cell clones that can express HYAL1 gene were obtained and can promote the invasion.

参考文献/References:

苏新良,任国胜,王小毅,等. 人乳腺癌细胞HYAL1基因真核表达载体的构建及其在MCF-7和ZR-75-30细胞中的表达[J].第三军医大学学报,2008,30(2):146-149.

相似文献/References:

[1]苏新良,任国胜,王小毅,等.HYAL1基因过表达对乳腺癌细胞迁移能力和血管生成能力及增殖的影响[J].第三军医大学学报,2008,30(15):1470.
 SU Xin-liang,REN Guo-sheng,WANG Xiao-yi,et al.Effects of HYAL1 gene overexpression on invasive, angiogenic and proliferative ability of breast cancer cells[J].J Third Mil Med Univ,2008,30(02):1470.
[2]王小毅,任国胜,石远,等.肝素对透明质酸酶不同表达水平的人乳腺癌细胞侵袭力的影响[J].第三军医大学学报,2002,24(10):0.[doi:10.16016/j.1000-5404.2002.10.042 ]
 WANG Xiao yi,REN Guo sheng,SHI Yuan,et al.[J].J Third Mil Med Univ,2002,24(02):0.[doi:10.16016/j.1000-5404.2002.10.042 ]

更新日期/Last Update: 2008-04-30