Construction and identification of CerbB-2 siRNA expression plasmid and its transfer into human colon cancer cell lines HT-29 / 南方医科大学学报
Journal of Southern Medical University
; (12): 1870-1873, 2009.
Article
in Zh
| WPRIM
| ID: wpr-336062
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a plasmid carrying small interfering RNA (siRNA) targeting human C-erbB2 gene (pGenesil- erbB2) and test its effect on Her-2 expression at the post-transcriptional level in human colon cancer cell lines HT-29 cells that highly express erbB2.</p><p><b>METHODS</b>A HT-29 cell line that highly expressed CerbB-2 was selected using immunohistochemical method. The double-stranded siRNA targeting human CerbB-2 cDNA and the negative control fragment were cloned into pGenesil-1 vector, and after identification and sequence analysis, the constructed pGenesil-erbB2 plasmid was transfected into the selected HT-29 cell line.</p><p><b>RESULTS</b>The pGenesil-erbB2 plasmid was successfully constructed and stably transferred into HT-29 cells. The transfection resulted in significant inhibition of Her-2 protein expression in the HT-29 cells, as shown by Western blotting.</p><p><b>CONCLUSION</b>The pGenesil-erbB plasmid we constructed can be stably transfected into HT-29 cells to inhibit the expression of Her-2 protein, and can be useful in further studies of increasing the radiosensitivity of HT-29 cell lines.</p>
Full text:
1
Database:
WPRIM
Main subject:
Plasmids
/
Molecular Sequence Data
/
Base Sequence
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Transfection
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Receptor, ErbB-2
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Genes, erbB-2
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HT29 Cells
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RNA, Small Interfering
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RNA Interference
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Genetics
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2009
Document type:
Article