【摘要】 目的:观察选择性环氧合酶-2(COX-2)抑制剂NS398对胰腺癌细胞周期及细胞周期相关蛋白p21Wafl/cipl、p27Kipl/pic2转录和表达的影响,探讨NS398的抗胰腺癌机制。方法:以NS398和前列腺素E2(PGE2)处理SW1990人胰腺癌细胞,分别采用MTT法检测细胞活力,酶联免疫分析法(ELISA)检测细胞内PGE2含量,流式细胞仪(FCM)检测细胞周期变化,并以半定量逆转录聚合酶链式反应(RT-PCR)和Western印迹检测COX-2及细胞周期相关蛋白p21Wafl/cipl、p27Kipl/pic2的mRNA和蛋白水平。结果:NS398抑制胰腺癌细胞生长,并呈剂量依赖性减少细胞内PGE2的生成;NS398诱导细胞周期相关蛋白p21Wafl/cipl和p27Kipl/pic2转录和表达的升高并诱导部分细胞阻滞在G0/G1期(较对照组升高11%);10 nmol/L的外源性PGE2可增加胰腺癌细胞的活力,但并不能拮抗NS398对细胞活力的抑制作用或细胞周期分布的改变,以及COX-2、p21Wafl/cipl、p27Kipl/pic2的转录和表达。结论:NS398可能通过增强p21Wafl/cipl和p27Kipl/pic2的表达而诱导细胞周期的阻滞,从而抑制胰腺癌细胞的生长活力。但NS398并非通过抑制COX-2的唯一机制,还可能存在其它非COX-2途径。更多还原

【Abstract】 AIM: To investigate the effects of selective cyclooxygenase-2(COX-2) inhibitor,NS398,on cell cycle and related cyclin-dependent kinase inhibitors p21waf1/cip1 and p27kip1/pic2 in human pancreatic carcinoma cells.METHODS: SW1990 human pancreatic carcinoma cells were treated with NS398(100 μmol/L),prostaglandin E2(PGE2,10 nmol/L) and their combination,respectively.The inhibitory effects of NS398 on SW1990 cell were detected by using MTT assay.The cell cycle was measured with flow cytometry.The level of intracellular PGE2 was determined with ELISA.The mRNA of p21waf1/cip1 and p27kip1/pic2 was detected by semi-quantitative RT-PCR.The expression of p21waf1/cip1 and p27kip1/pic2 protein was detected by Western blotting analysis.RESULTS: NS398 inhibited the growth of SW1990 cell and decreased level of intracellular PGE2 in a dose-dependent manner.NS398 caused cell accumulation in G0/G1 phase that was 11% higher than control.The mRNA and protein of p21waf1/cip1 and p27kip1/pic2 were up-regulated by NS398.PGE2 stimulated cell growth,but factorial experiment showed that 10 nmol/L of PGE2 could not antagonize inhibitory effects of cell growth,cell cycle distribution,transcription and expression of COX-2,p21waf1/cip1 and p27kip1/pic2 induced by NS398 in SW1990 cells.CONCLUSION: The results suggest that selective COX-2 inhibitor,NS398,can induce inhibitory effects of cell growth and G0/G1 cell cycle arrest in SW1990 cells by up-regulation of p21waf1/cip1 and p27kip1/pic2 and may be not dependent of PGE2 pathway.更多还原