Retrovirus-mediated SHP-1 gene expression in human breast cancer MDA-MB-231 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 902-905, 2009.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-268815
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a retrovirus-mediated expression system carrying human SHP-1 gene to transfer SHP-1 gene in human breast cancer MDA-MB-231 cells.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>A 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.</p><p><b>CONCLUSION</b>The 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.</p>
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Retroviridae
/
Recombinant Proteins
/
Breast Neoplasms
/
Transfection
/
Cloning, Molecular
/
Reverse Transcriptase Polymerase Chain Reaction
/
Cell Line, Tumor
/
Protein Tyrosine Phosphatase, Non-Receptor Type 6
/
Genetic Vectors
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2009
Document type:
Article