[Cloning and expression of the HER2 gene in MCF-7 cells].
OBJECTIVE
To clone human epidermal growth factor receptor (HER2) gene and establish a MCF-7 cell line with stable co-expression of HER2 and estrogen receptor (ER).


METHODS
The full-length HER2 gene fragment was amplified by RT-PCR from the total RNA extracted from human breast cancer cell line SKBR-3 which over-expressed HER2 protein. The gene fragment was then inserted into the vector pcDNA3.1 to construct the recombinant expression plasmid pcDNA3.1-HER2, which was identified by sequence analysis and transfected into ER-expressing MCF-7 cells via lipofectamine. The positive cell clones were obtained after G418 selection. The MCF-7 cells transfected with HER2-cDNA was assayed by immunocytochemistry, flow cytometry and Western blotting.


RESULTS
DNA sequencing results showed that HER2 gene was exactly consistent with the sequence reported in GenBank. The MCF-7 cells transfected with HER2 could highly express HER2 protein as shown by immunocytochemistry, flow cytometry and Western blotting.


CONCLUSIONS
HER2 gene has been cloned successfully and the MCF-7 cell line co-expressing HER2 and ER is obtained.
Li
Rong
R
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Zheng
Hang
H
Luo
Rong-cheng
RC
D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005260:Female; D018734:Genes, erbB-2 / Q000235:genetics*; D006801:Humans; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D018719:Erb-b2 Receptor Tyrosine Kinases