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目的 :探讨环氧化酶(cyclooxygenase-2,COX-2)抑制剂塞来昔布对人红白血病HEL细胞增殖、凋亡及迁移的影响及其作用机制。方法:不同浓度的塞来昔布处理HEL细胞,CCK-8法检测细胞增殖抑制率;Hoechst33342荧光染色检测细胞凋亡;Transwell小室检测细胞迁移率;RT-PCR检测COX-2及JAK2 mRNA水平;Western blot检测COX-2及p-JAK2蛋白表达。结果:塞来昔布能够时间和剂量依赖性抑制HEL细胞增殖,不同浓度(25、75、125μmol/L)的塞来昔布在48 h时对细胞生长抑制率分别为(9.96±0.82)%、(18.46±2.01)%、(21.36±2.48)%(P<0.05);Hoechst33342凋亡细胞染色显示125μmol/L塞来昔布处理HEL细胞后,凋亡细胞明显增多;细胞迁移实验结果显示75μmol/L塞来昔布处理细胞24 h后漏出细胞为(22.13±7.51)个,明显低于对照组(77.89±6.94)个,P<0.05;RT-PCR结果显示不同浓度塞来昔布处理HEL细胞48 h后COX-2 mRNA呈剂量依赖性减低,而对JAK2 m RNA无明显影响;Western blot结果显示塞来昔布处理HEL细胞COX-2蛋白表达明显减低(P<0.05),而对p-JAK2无明显影响。结论:塞来昔布能够抑制HEL细胞增殖,可能与抑制COX-2表达有关,而对JAK2信号通路无明显影响。
Objective: To investigate the effect of cyclooxygenase-2 (COX-2) inhibitor celecoxib on proliferation, apoptosis and migration of human erythroleukemia HEL cells and its mechanism. METHODS: HEL cells were treated with different concentrations of celecoxib, the cell proliferation inhibition rate was determined by CCK-8 assay, the apoptosis was detected by Hoechst33342 staining, the cell migration rate was detected by Transwell chamber, the mRNA expression of COX-2 and JAK2 was detected by RT- Western blot detection of COX-2 and p-JAK2 protein expression. RESULTS: Celecoxib inhibited the proliferation of HEL cells in a time and dose-dependent manner. The inhibitory rates of celecoxib at different concentrations (25, 75 and 125μmol / L) were (9.96 ± 0.82)% at 48 h , (18.46 ± 2.01)% and (21.36 ± 2.48)%, respectively (P <0.05). The apoptosis of Hoechst33342 cells showed that the apoptotic cells were significantly increased after treated with 125μmol / L Celecoxib. The results of cell migration showed that 75μmol / L celecoxib treatment cells (22.13 ± 7.51), significantly lower than the control group (77.89 ± 6.94), P <0.05; RT-PCR results showed that different concentrations of celecoxib treatment of HEL After 48 h, COX-2 mRNA decreased in a dose-dependent manner, but had no effect on JAK2 m RNA. The Western blot results showed that the COX-2 protein expression in HEL cells decreased significantly (P <0.05) -JAK2 No significant effect. Conclusion: Celecoxib can inhibit the proliferation of HEL cells, which may be related to the inhibition of COX-2 expression, but not to JAK2 signaling pathway.