Proapoptotic and anti proliferative potential of selective cyclooxygenase-2 inhibitors in human liver tumor cells
Proapoptotic and anti proliferative potential of selective cyclooxygenase-2 inhibitors in human liver tumor cells
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DOI：
10.1053/jhep.2002.36125
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发表时间：
2002-10-01
期刊：
影响因子：
13.5
通讯作者：
Schirmacher, P
中科院分区：
文献类型：
--
作者：
Kern, MA;Schubert, D;Schirmacher, P
近年来的研究表明，环氧合酶-2（考克斯-2）在包括肝细胞癌（HCC）在内的多种人类恶性肿瘤中的水平升高，但到目前为止，考克斯-2是否有助于恶性生长以及抑制考克斯-2功能是否改变肝肿瘤的恶性潜能尚不清楚。采用北方杂交和Western免疫印迹法检测了4种肝癌细胞系（Hep 3B、HuH-7、Hep G2、Sk-hep 1）中考克斯-1和考克斯-2的表达。非选择性抑制剂舒林酸硫化物和考克斯-2选择性抑制剂SC-58635和美洛昔康的功能效应通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四唑溴化物（MTT）-测定和BrdU摄取、形态学和凋亡的TUNEL分析来检查。通过Western免疫印迹分析凋亡调节蛋白。考克斯-1和考克斯-2的表达在所有测试的肝肿瘤细胞系中是可证实的。舒林酸硫化物（50 - 400 mumol/L）、SC-58635（6.25 - 400 mumol/L）和美洛昔康（6.25 - 400 mumol/L）导致细胞数量显著减少，最高达80%（P <0.05），呈时间和剂量依赖性。在等摩尔浓度下，当选择性阻断考克斯-2时，效果更明显。抑制考克斯-2可诱导细胞凋亡并减少肿瘤细胞增殖。SC-58635（50 mumol/L）抑制考克斯-2后的细胞凋亡与BCL-2、BAX以及AKT/PKB和BAD的磷酸化状态无关，但与caspase-9、caspase-3和caspase-6的活化相关。总之，选择性抑制考克斯-2导致显着的生长抑制人肝肿瘤细胞的基础上，诱导细胞凋亡和增殖的抑制，因此，可能提供治疗和预防人类肝癌的潜力。
Recent studies have shown increased levels of cyclooxygenase-2 (COX-2) in a variety of human malignancies, including hepatocellular carcinoma (HCC), but so far it is unknown whether COX-2 contributes to the malignant growth and whether inhibition of COX-2 function modifies the malignant potential of liver tumors. COX-1 and COX-2 expression was determined in 4 liver tumor cell lines (Hep 3B, HuH-7, Hep G2, Sk-hep1) by Northern hybridization and Western immunoblot. The functional effects of the nonselective inhibitor sulindac sulfide and the COX-2 selective inhibitors SC-58635 and meloxicam were examined by 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide (MTT)-assays and BrdU uptake, morphology, and TUNEL analysis of apoptosis. Apoptosis regulating proteins were analyzed by Western immunoblot. COX-1 and COX-2 expression was demonstrable in all tested liver tumor cell lines. Sulindac sulfide (50 to 400 mumol/L), SC-58635 (6,25 to 400 mumol/L), and meloxicam (6.25 to 400 mumol/L) led to a significant time- and dose-dependent reduction of cell numbers of up to 80% (P < .05). At equimolar concentrations the effect was more pronounced when COX-2 was selectively blocked. COX-2 inhibition induced apoptosis and reduced tumor cell proliferation. Apoptosis after COX-2 inhibition with SC-58635 (50 mumol/L) was independent of BCL-2, BAX, and the phosphorylation status of AKT/PKB and BAD, but correlated with activation of caspase-9, caspase-3, and caspase-6. In conclusion, selective inhibition of COX-2 leads to a marked growth inhibition of human liver tumor cells, based on the induction of apoptosis and inhibition of proliferation and, thus, may offer therapeutic and preventive potential in human hepatocarcinogenesis.