[Effect of down regulation of laminin receptor on urokinase-type plasminogen activator expression in human bile duct carcinoma cells].
OBJECTIVE
To investigate the effect of antisense oligonucleotide (AS-OD) of laminin receptor (LNR) on urokinase-type plasminogen activator (u-PA) expression in human bile duct carcinoma cells.


METHODS
Bile duct carcinoma cells of QBC939 line were cultured. Compounds of lipofectAMINE and AS-OD phosphorothioate of LNR of the concentrations of 3, 6, and 12 micromol/L respectively were prepared and then transfected into the QBC939 cells. Sense oligodeoxynucleotide (S-OD) of LNR and blank vector were used as controls. After culture of 48 hours, flow cytometry was used to detect the protein expression of LNR and the u-PA mRNA expression was detected by reverse-transcriptase polymerase chain reaction (RT-PCR). Cell invasion was examined by culture plate with Transwell microporous film before and after transfection.


RESULTS
Flow cytometry showed down-regulation of LNR protein expression in the cells transfected with AS-OD dose-dependently. The LNR protein expression of the cells transfected with AS-OD of the concentration of 6 micromol/L was down-regulated by over 25% in comparison with those of the blank vector group and S-OD group (both P < 0.05). RT-PCR showed that the u-PA mRNA expression of the AS-OD 6 micromol/L group was down-regulated by 30% in comparison with the 2 control groups. The number of migrating cells in the AS-OD group was 78 +/- 6, significantly lower than those in the S-OD group (105 +/- 11), and blank vector group (101 +/- 7), both P < 0.05.


CONCLUSIONS
The expression of laminin receptor in the bile duct carcinoma cells is related with the expression of u-PA gene expression. Inhibition of LNR expression decreases the u-PA gene expression, suggesting that LNR plays an important role in the invasion and metastasis of bile duct carcinoma.
Wang
Shu-guang
SG
Institute of Hepatobiliary Surgery, People's Liberation Army Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Chen
Chang-hong
CH
Li
Da-jiang
DJ
Li
Kun
K
D001650:Bile Duct Neoplasms / Q000378:metabolism*; D015536:Down-Regulation; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D017474:Receptors, Laminin / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis* / Q000235:genetics
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D017474:Receptors, Laminin; D014568:Urokinase-Type Plasminogen Activator