摘要
目的检测体外培养的人肝细胞癌HepG2,Hep3B细胞和正常肝细胞系QSG7701细胞内环氧酶-2(cyclooxygenase-2,COX-2)的表达,观察环氧酶抑制剂双氯芬酸(diclofenac)对上述细胞增殖的作用,并检测前列腺素E2(PGE2)对双氯芬酸作用的影响,以探讨双氯芬酸抗增殖作用与细胞内COX-2的关系。方法采用半定量逆转录聚合酶链反应(RT-PCR)检测COX-2 mRNA的表达水平,免疫印迹技术检测细胞内蛋白水平,cellcounting kit-8(CCK-8)细胞计数法测定细胞增殖活性。结果肝细胞癌HepG2和Hep3B细胞均高表达COX-2 mRNA和COX-2蛋白,正常肝细胞系QSG7701微弱表达COX-2。10～200μmol·L-1的双氯芬酸浓度依赖性地抑制HepG2和Hep3B细胞的增殖(P<0.01),作用72h的半数抑制浓度(IC50)分别为76.41和35.21μmol·L-1。双氯芬酸对QSG7701的抑制作用较弱,IC50值为170.23μmol·L-1。20μmol·L-1 PGE2与50μmol·L-1双氯芬酸共同孵育能显著削弱后者抗HepG2和Hep3B细胞增殖的作用(P<0.01)。结论双氯芬酸通过抑制COX-2活性和PGE2的生成,特异性地抑制表达COX-2的肝癌细胞的增殖。
Abstract
OBJECTIVE To investigate the cyclooxygenase-2(COX-2) mRNA and protein expression of cultured hepatoma cell lines HepG2,Hep3B and human hepatocellular cell line QSG7701,and detect the effects of the cyclooxygenase inhibitor diclofenac on proliferation of those cells,and determine the effect of prostaglandin E2(PGE2) on the action of diclofenac. METHODS COX-2 mRNA and protein expression were determined by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) and immunoblotting respectively.Cell viability was analyzed by cell counting kit-8(CCK-8) assay. RESULTS COX-2 mRNA and protein were highly expressed in HepG2 and Hep3B cells,but hardly in QSG7701 cells. Diclofenac (10～200 μmol·L-1) exposure for 72 h significantly suppressed HepG2 and Hep3B cells growth in a concentration-dependened manner (P<0.01),with the 50 percent inhibitory concentration (IC50) of 76.41 and 35.21 μmol·L-1 respectively.A much weaker anti-proliferation effect on QSG7701 was shown,with the IC50 of 170.23 μmol·L-1.20 μmol·L-1 PGE2 significantly prevented the antiproliferation effect of 50 μmol·L-1 diclofenac on HepG2 and Hep3B cells(P<0.01). CONCLUSION Diclofenac specifically suppresses proliferation of COX-2 expressing hepatoma cells through inhibition of COX-2 activity and PGE2 production.
关键词
双氯芬酸 /
肝细胞癌 /
细胞增殖 /
环氧合酶-2 /
前列腺素E2
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Key words
diclofenac /
hepatoma /
cell proliferation /
cyclooxygenase-2 /
prostaglandin E2
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陈娜娜;吴曙光.
双氯芬酸选择性抑制表达环氧合酶-2的肝癌细胞的增殖[J]. 中国药学杂志, 2007, 42(15): 1152-1154
CHEN N-n;WU Shu-gung.
Inhibtion of Diclofenac on Proliferation of Hepatoma Cells Expressing Cyclooxygenase-2 [J]. Chinese Pharmaceutical Journal, 2007, 42(15): 1152-1154
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脚注
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