塞来昔布对胶质瘤细胞生物学特性的影响

The effect of celecoxib on the biological characteristics of glioma cells

目的观察塞来昔布对胶质瘤细胞增殖、凋亡、迁移及侵袭能力影响。方法噻唑蓝（MTT）法检测胶质瘤细胞U251、T98G、U87经过0、20、40、60 μmol/L的塞来昔布药物作用24、48、72、96 h后的细胞增殖情况，同时检测正常的星型细胞HA经塞来昔布作用后的细胞增殖。流式细胞仪检测40 μmol/L的塞来昔布作用48 h后的胶质瘤细胞U251凋亡。细胞划痕实验检测40 μmol/L的塞来昔布作用48 h后的胶质瘤细胞U251迁移。Transwell小室检测40 μmol/L的塞来昔布作用16 h后的胶质瘤细胞U251侵袭情况。Western blot检测细胞中环氧化酶-2（COX-2）、B淋巴细胞瘤-2（bcl-2）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）、基质金属蛋白酶（MMP）-9、MMP-2水平。结果塞来昔布对胶质瘤细胞增殖具有抑制作用，对正常的星型细胞无明显的抑制作用。塞来昔布对胶质瘤细胞增殖抑制作用随着作用时间的增加而增加，随着药物作用浓度的增加而增加。经40 μmol/L的塞来昔布药物作用后的胶质瘤细胞凋亡率高于对照组[（3.17±0.16）%比（16.08±1.57）%，t＝14.169，P＝0.000]。经40 μmol/L的塞来昔布作用后的胶质瘤细胞U251的迁移率明显低于对照组[（100.00±5.26）%比（41.28±5.84）%，t＝12.940，P＝0.000]。塞来昔布作用后的胶质瘤细胞U251的侵袭能力明显低于对照组[（229.5±11.36）个比（97.32±7.95）个，t＝16.512，P＝0.000]。塞来昔布作用后的胶质瘤细胞中COX-2、bcl-2、MMP-9、MMP-2的水平较对照组比较明显下降，塞来昔布作用后的胶质瘤细胞中bax的水平较对照组比较明显升高（t1＝19.696，P1＝0.000；t2＝17.774，P2＝0.000；t3＝12.970，P3＝0.000；t4＝10.569，P4＝0.001）。结论塞来昔布可以抑制胶质瘤细胞增殖、迁移、侵袭能力，促进胶质瘤细胞凋亡，其作用机制与COX-2、bax、bcl-2、MMP-9、MMP-2有关。

Objective To investigate the effect of celecoxib on glioma cell proliferation, apoptosis, migration and invasion ability.Methods Methyl thiazol tetrazolium （MTT） assay was used to detect glial tumor cell proliferation of U251, T98G, U87 after treatment with 0, 20, 40 or 60 μmol/L celecoxib for 24, 48, 72 or 96 h. Flow cytometry was used to detect cell apoptosis after treatment with 40 μmol/L celecoxib for 48 h. Cell scratch test was used to detect cell migration after treatment with 40 μmol/L celecoxib for 48 h. Transwell was used to detect cell invasion after treatment with 40 μmol/L celecoxib for 48 h. The expression levels of cyclooxygenase-2 （COX-2）, B cell lymphoma/leukemia-2 （bcl-2）, B cell lymphoma/leukemia-2 associated X protein （bax）, matrix metalloproteinase （MMP）-9, MMP-2 in cells was detected by Western blotting.Results Celecoxib can inhibit the proliferation of glioma cells and there is no obvious inhibitory effect on normal astrocytoma. The inhibitory effect of Celecoxib on glioma cell proliferation was depended on the reaction time and drug concentration. The apoptosis rate of glioma cell after 40μmol/L celecoxib was higher than the control group, and the difference was significant [（3.17±0.16）% vs. （16.08±1.57）%, （3.17±0.16）% vs. （16.08±1.57）%, t=14.169, P=0.000]. The migration rate of glioma cell after treatment with 40 μmol/L celecoxib was lower than the control group, the difference was significant . The invasion ability of glioma cell after 40 μmol/L celecoxib was lower than the control group, the difference was significant [（100.00±5.26）% vs. （41.28±5.84）%, t=12.940, P=0.000]. Glioma cells after celecoxib treatment in COX-2, bcl-2, MMP-9, MMP-2 was significantly decreased than control group [（229.5±11.36） vs. （97.32±7.95）, t=16.512, P=0.000]. The bax protein of Glioma cells after celecoxib treatmentwas significantly higher than control group （t1=19.696, P1=0.000; t2=17.774, P2=0.000; t3=12.970, P3=0.000; t4=10.569, P4=0.001）.Conclusion Celecoxib can inhibit glioma cell proliferation, migration and invasion ability, and promote the apoptosis of glioma cells, the mechanism of which was related to COX-2, bcl-2, bax, MMP-9, MMP-2.