Retrovirus-mediated SHP-1 gene expression in human breast cancer MDA-MB-231 cells.
- Author:
Ji-zhen LIANG
1
;
Rong LI
;
Jun-yi ZHANG
;
Hang ZHENG
;
Rong-cheng LUO
Author Information
- Publication Type:Journal Article
- MeSH: Breast Neoplasms; genetics; metabolism; pathology; Cell Line, Tumor; Cloning, Molecular; Genetic Vectors; genetics; Humans; Protein Tyrosine Phosphatase, Non-Receptor Type 6; biosynthesis; genetics; Recombinant Proteins; biosynthesis; genetics; Retroviridae; genetics; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Transfection
- From: Journal of Southern Medical University 2009;29(5):902-905
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a retrovirus-mediated expression system carrying human SHP-1 gene to transfer SHP-1 gene in human breast cancer MDA-MB-231 cells.
METHODSThe full-length SHP-1 gene fragment was amplified by RT-PCR from the total RNA extracted from human breast cancer cell line MCF-7 over-expressing SHP-1 protein. The gene fragment was inserted into the vector pLNCX2 to construct the recombinant retroviral plasmid, which was transfected into the packaging cell PT67 via Lipofectamine2000. A cell line stably producing the virus was selected with G418. MDA-MB-231 cells was infected with the virus, and the expression of SHP-1 gene in the positive cell clone was detected with Western blotting.
RESULTSA 1.8 kb cDNA fragment of SHP-1 gene was obtained from MCF-7 cells and successfully inserted into the pLNCX2. A stable cell clone PT67/SHP-1 and virus supernatant were obtained. Expression of SHP-1 protein was detected in the cells infected with the virus.
CONCLUSIONThe recombinant retroviral vector carrying SHP-1 gene has been successfully constructed and MDA- MB-231/SHP-1 cell line expressing SHP-1 has been obtained to allow further functional study of SHP-1 in breast cancer.