COX-2抑制剂对人红白血病HEL细胞增殖、凋亡及迁移的影响
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河北省重点研发科技项目(162777120D);保定市科学技术研究与发展指导计划(12ZF105)


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

    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(25-75-125 μmol/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125 μmol/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75 μmol/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 mRNA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。

    Abstract:

    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 mRNA 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 with 75 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 mRNA was dependently decreased, while no significant effect on the JAK2 mRNA 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.

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赵亚玲,刘贵敏,梁文同,成志勇,谢旭磊,谷 蕾,李 琳,颜晓燕. COX-2抑制剂对人红白血病HEL细胞增殖、凋亡及迁移的影响[J].南京医科大学学报(自然科学版),2016,(4):435-439

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  • 收稿日期:2015-07-26
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  • 在线发布日期: 2016-05-11
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