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
Dans Chinois
| WPRIM
| ID: wpr-268815
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>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Anatomopathologie
/
Retroviridae
/
Protéines recombinantes
/
Tumeurs du sein
/
Transfection
/
Clonage moléculaire
/
RT-PCR
/
Lignée cellulaire tumorale
/
Protein Tyrosine Phosphatase, Non-Receptor Type 6
/
Vecteurs génétiques
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2009
Type:
Article
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