Construction of eukaryotic expression vector of human HYAL1 gene and its expression in MCF-7 and ZR-75-30 cells
- VernacularTitle: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
- Author:
Xinliang SU
;
Guosheng REN
;
Xiaoyi WANG
;
Jinxiang TAN
- Publication Type:Journal Article
- Keywords:
hyaluronidase;
HYAL1;
eukaryotic expression;
breast cancer cell;
invasion
- From:Journal of Third Military Medical University
1984;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo construct the eukaryotic expression vector of human HYAL1 gene and obtain MCF-7 and ZR-75-30 cell clones expressing HYAL1 gene stably.MethodsThe 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.ResultsA 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.ConclusionThe 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.
