COX-2抑制剂对人红白血病HEL细胞增殖、凋亡及迁移的影响

Effects of COX-2 inhibitor on proliferation, apoptosis and migration of human erythroleukemia HEL cells

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 COX-2 inhibitor effect of celecoxib on proliferation, apoptosis and migration of human erythroleukemia HEL cells and its mechanism. Methods： The human erythroleukemia HEL cells were treated with different concentrations of celecoxib. The cell proliferation inhibition rate was calculated by CCK-8 test; the apoptosis rate was detected by Hoechst33342 fluorescent staining; cell migration ability was tested by transwell chambers. The expression levels of COX-2 and JAK2 m RNA were detected by Real-time PCR; the protein expression levels of COX2 and p-JAK2 were detected by Western blotting assay.Results： Celecoxib time and dose dependently inhibited the proliferation of HEL cells, the cell growth inhibition rates were（9.96 ± 0.82）%,（18.46 ± 2.01）% and（21.36 ± 2.48）%, respectively（P〈0.05）, after treated with different concentrations（25,75,and 125 μmol / L,respectively） of celecoxib in HEL cells after 48 h. Hoechst33342 fluorescent staining showed that apoptosis increased significantly after treatment of 125 μmol / L celecoxib in HEL cells after 48 h; cell migration ability showed that the leakage after treated with75 μmol / L celecoxib in HEL cells after 24 h was 22.13 ± 7.51, which was significantly lower than that of the control group（77.89 ±6.94, P〈0.05）; RT-PCR results showed that COX-2 m RNA was dependently decreased, while no significant effect on the JAK2 m RNA after treated with different concentrations of celecoxib; Western blotting assay showed that the expression of COX-2 protein in the experimental group was significantly lower than that in the control group（P〈0.05）, but no obvious effect was found on p-JAK2 expression. Conclusion： Proliferation of HEL cells can be inhibited by celecoxib. It may be related to the inhibition of the expression of COX-2, but had no obvious effect on JAK2 signaling pathway.