双氯芬酸抑制肝癌细胞增殖和促进环氧合酶-2 mRNA作用的表达

Diclofenac suppresses hepatoma cell proliferation and promotes cyclooxygenase-2 mRNA expression

目的观察环氧合酶抑制剂双氯芬酸钠对体外培养的肝癌细胞HepG2、Hep3B和正常肝细胞系QSG-7701增殖的作用以及药物对环氧合酶-2(COX-2)mRNA表达的影响,探讨COX-2与肝癌细胞增殖的关系。方法采用CellCountingKit-8(CCK-8)细胞计数法测定药物作用24、48、72h后细胞增殖活性,半定量逆转录聚合酶链反应检测各种细胞系COX-2mRNA的表达水平以及药物作用对基因表达的影响。结果双氯芬酸在10-200μmol/L浓度依赖性地抑制HepG2和Hep3B细胞的增殖,50μmol/L作用48h后抑制率分别为40.47%和54.49%,半数抑制浓度分别为70.54和48.39μmol/L。双氯芬酸对正常肝细胞株QSG-7701的抑制作用较弱,作用48h的半数抑制浓度为189.91μmol/。HepG2、Hep3B细胞均表达COX-2mRNA,QSG-7701细胞几乎不表达COX-2mRNA。双氯芬酸和化疗药5-氟尿嘧啶均可增强HepG2和Hep3B细胞COX-2mRNA的表达。结论双氯芬酸特异性地抑制表达COX-2的肝癌细胞HepG2和Hep3B的增殖,并且诱导COX-2mRNA表达上调,提示COX-2在肝癌细胞增殖中具有重要意义。

Objective To investigate the effects of cyclooxygenase inhibitor diclofenac on the proliferation and cyclooxygenase-2 （COX-2） mRNA expression of cultured hepatocellular carcinoma cell lines HepG2, Hep3B and human hepatocellular cell line QSG-7701. Methods After exposure to diclofenac at various concentrations （10-200 μmol/L） for 24, 48 and 72 h, the cell proliferation was analyzed by Cell Counting Kit-8 （CCK-8） assay and mRNA expression determined by semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR）. Results Diclofenac exposure for 24, 48 and 72 h significantly inhibited HepG2 and Hep3B cell proliferation in a concentration-dependent manner, with inhibition rate of 40.47% and 54.49% after 48 h exposure to 50 μmol/L diclofenac and IC50 of 70.54 and 48.39 μmol/L, respectively. A much weaker antiproliferative effect on QSG-7701 cells was shown, with IC50 of 189.91 μmol/L after 48-hour exposure to diclofenac. RT-PCR detected COX-2 mRNA in HepG2 and Hep3B cells, but hardly in QSG-7701 cells. Treatment with diclofenac or 5-Fu resulted in elevated COX-2 mRNA expression both in HepG2 and Hep3B cells. Conclusions Diclofenac can specifically inhibit the proliferation of COX-2-expressing HepG2 and Hep3B cells, and induce up-regulation of COX-2 mRNA expression, which indicates the important role of COX-2 in the proliferation of hepatoma cells.