The selective cyclooxygenase-2 inhibitor nimesulide induces apoptosis in pancreatic cancer cells independent of COX-2
The selective cyclooxygenase-2 inhibitor nimesulide induces apoptosis in pancreatic cancer cells independent of COX-2
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DOI：
10.1097/00006676-200301000-00007
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发表时间：
2003-01-01
期刊：
影响因子：
2.9
通讯作者：
Hines, OJ
中科院分区：
文献类型：
--
作者：
Eibl, G;Reber, HA;Hines, OJ
简介：选择性环氧合酶-2（考克斯-2）抑制可降低许多癌细胞系的生长，这种作用可能通过诱导细胞凋亡来介导。本研究旨在探讨考克斯-2抑制剂尼美舒利对人胰腺癌细胞生长的影响。方法学：逆转录聚合酶链反应分析，北方印迹，免疫印迹法被用来证明考克斯-2 mRNA和蛋白在两个胰腺癌细胞系：BxPC-3和MIA PaCa-2。尼美舒利对细胞生长的影响通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑测定和细胞计数来评估。通过流式细胞术分析、DNA梯状电泳和评估漂浮细胞/附着细胞比率来确定细胞凋亡。通过测量胱天蛋白酶-3细胞活性和通过测定两种胱天蛋白酶抑制剂对细胞活力的影响来评价胱天蛋白酶-3活化。结果：两种细胞株均检测到考克斯-2 mRNA的表达。免疫印迹证实考克斯-2蛋白仅在BxPC-3细胞系中表达。尼美舒利在两种细胞系中均降低细胞活力并抑制细胞生长。用100 μ mol/L尼美舒利孵育增加了两种细胞系中凋亡细胞的比例和漂浮细胞/贴壁细胞的比例。在与尼美舒利孵育96小时后观察到DNA梯状条带。在96小时内，caspase-3活性没有增加。结论：选择性考克斯-2抑制剂尼美舒利对胰腺癌细胞具有抗有丝分裂作用，其作用不依赖于考克斯-2的表达。这种作用部分是通过诱导细胞凋亡介导的。
Introduction: Selective cyclooxygenase-2 (COX-2) inhibition reduces the growth of many cancer cell lines, and this effect may be mediated through induction of apoptosis. This study was designed to investigate the effects of the COX-2 inhibitor nimesulide on the growth of human pancreatic cancer cells. Methodology: Reverse transcriptase-polymerase chain reaction analysis, northern blotting, and immunoblotting were used to demonstrate COX-2 mRNA and protein in two pancreatic cancer cell lines: BxPC-3 and MIA PaCa-2. The effect of nimesulide on cell growth was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and cell count. Apoptosis was determined by FACS analysis, DNA laddering, and assessment of the floating cell/attached cell ratio. Caspase-3 activation was evaluated by measuring caspase-3 cellular activity and by determining the effects of two caspase inhibitors on cell viability. Results: COX-2 mRNA was detected in both cell lines. Immunoblotting confirmed COX-2 protein expression in only the BxPC-3 cell line. Nimesulide decreased cell viability and inhibited cell growth in both cell lines. Incubation with 100 mumol/L nimesulide increased the fraction of apoptotic cells and the floating cell/attached cell ratio in both cell lines. DNA laddering was observed after incubation with nimesulide for 96 hours. There was no increase in caspase-3 activity within 96 hours. Conclusion: The selective COX-2 inhibitor nimesulide is antimitogenic in pancreatic cancer cells, which is independent of COX-2 expression. This effect in part is mediated by the induction of apoptosis.