Effects of cyclooxygenase-2 antisense vector on proliferation of human cholangiocarcinoma cells.
OBJECTIVE
To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis.


METHODS
QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry.


RESULTS
RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P < 0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P > 0.05).


CONCLUSIONS
Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.
Wu
Gao-Song
GS
Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030. wugs@tjh.tjmu.edu.cn
Zou
Sheng-Quan
SQ
Wu
Xiao-Yong
XY
Qiu
Fa-Zu
FZ
D017209:Apoptosis; D001650:Bile Duct Neoplasms / Q000378:metabolism / Q000473:pathology*; D001653:Bile Ducts, Intrahepatic; D002453:Cell Cycle; D002455:Cell Division; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000378:metabolism / Q000473:pathology*; D051546:Cyclooxygenase 2; D016373:DNA, Antisense / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D008565:Membrane Proteins; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection
D016373:DNA, Antisense; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases