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Effects of cyclooxygenase-2 antisense vector on proliferation of human cholangiocarcinoma cells

Wu, G-Song.; Zou, S-Quan.; Wu, X-Yong.; Qiu, F-Zu.

Chinese Medical Sciences Journal 19(2): 89-92

2004

ISSN/ISBN: 1001-9294
PMID: 15250241
Document Number: 579299
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(TM) 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). Conclusion Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

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