[Establishment of a colorectal cancer cell line with PRL-3 and CDH22 gene knock-down by lentivirus-mediated RNA interference].
OBJECTIVE
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.


METHODS
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.


CONCLUSIONS
A human colorectal cancer cell line with stable PRL-3 and CDH22 gene knock-down has been established.
Yao
Juan
J
Department of Pathology, Nanfang Hospital and School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Ding
Yan-qing
YQ
Zhou
Jun
J
Liu
Yu-hong
YH
Li
Jian-ming
JM
D015820:Cadherins / Q000235:genetics*; D045744:Cell Line, Tumor; D002999:Clone Cells / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000235:genetics / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques / Q000379:methods*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D017027:Protein Tyrosine Phosphatases / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D015820:Cadherins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; C099417:CDH22 protein, human; C484730:PTP4A3 protein, human; D017027:Protein Tyrosine Phosphatases