Construction of cyclin D1 recombinant plasmids and its expression in human nasopharyngeal carcinoma cells / 南方医科大学学报
Journal of Southern Medical University
;
(12): 202-205, 2010.
Article
Dans Chinois
| WPRIM
| ID: wpr-269593
ABSTRACT
<p><b>OBJECTIVE</b>To construct the eukaryotic expression vectors of human cyclin D1 gene and express them in poorly differentiated nasopharyngeal carcinoma cells (CNE-2Z cells).</p><p><b>METHODS</b>The full-length cyclin D1 was cloned from CNE-2Z cells by RT-PCR. The cDNA fragments were inserted into pIRES2-EGFP plasmids and pEGFP-C2 plasmids and confirmed by restriction enzyme digestion, PCR and sequencing. The recombinant vectors were transfected into CNE-2Z cells via Lipofectamine 2000, and the expression of cyclin D1 in the cells was examined by immunofluorescence and Western blotting.</p><p><b>RESULTS</b>Agarose gel electrophoresis showed a 918 bp band of the RT-PCR products, which matched the expected size. Restriction enzyme digestion, PCR and sequencing demonstrated successful construction of the recombinant vectors. CNE-2Z cells transfected with the recombinant vectors expressed cyclin D1 protein or cyclin D1-GFP protein as were verified by immunofluorescence and Western blotting.</p><p><b>CONCLUSION</b>We have cloned cyclin D1 gene and constructed its eukaryotic expression vectors that can be expressed in nasopharyngeal carcinoma cells, which may facilitate the study of the role of cyclin D1 in the development of nasopharyngeal carcinoma.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Anatomopathologie
/
Plasmides
/
Protéines recombinantes
/
Transfection
/
Tumeurs du rhinopharynx
/
Clonage moléculaire
/
Cycline D1
/
Lignée cellulaire tumorale
/
Protéines à fluorescence verte
/
Vecteurs génétiques
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2010
Type:
Article
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