Establishment of a colorectal cancer cell line with PRL-3 and CDH22 gene knock-down by lentivirus-mediated RNA interference / 南方医科大学学报
Journal of Southern Medical University
;
(12): 593-597, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-233733
ABSTRACT
<p><b>OBJECTIVE</b>To establish a colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down for investigating the role of PRL-3 and CDH22 genes in the carcinogenesis and metastasis of colorectal cancer.</p><p><b>METHODS</b>A recombinant lentiviral vector targeting CDH22 gene was obtained using the pENTRTM/U6 construct and pLenti6/BLOCK-iT (TM)-DEST vector. The recombinant lentivirus was harvested from 293FT cells cotransfected with the optimized ViraPower(TM) Packaging Mix and the pLenti6/BLOCK-iT(TM)-DEST expression construct. SW480/PRL-3- cells were infected with the recombinant lentivirus targeting CDH22, and SW480 cells with stable PRL-3 and CDH22 knock-down were screened by blasticidin selection. PRL-3 expression in the cells was determined by real-time RT-PCR. RESULTS The titer of the lentivirus for the second infection was 8 x 10(5) U/ml. Seventeen positive clones were selected, among which the Clone 1 exhibited substantially down-regulated CDH22 and PRL-3 mRNA expressions.</p><p><b>CONCLUSIONS</b>A human colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down has been established.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
RNA, Messenger
/
Colorectal Neoplasms
/
Cadherins
/
Gene Expression Regulation, Neoplastic
/
Protein Tyrosine Phosphatases
/
Clone Cells
/
Lentivirus
/
RNA Interference
/
Cell Line, Tumor
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2009
Type:
Article
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