Celecoxib对胆囊癌细胞GBC-SD增殖的抑制作用

Cyclooxygenase-2 specific inhibitor celecoxib inhibits proliferation and induces apoptosis in human carcinoma of gallbladder cell GBC-SD

目的 研究Celecoxib对人胆囊癌细胞GBC SD生长和凋亡的影响及其作用机制。方法 采用免疫细胞化学SABC法检测GBC SD环氧合酶 2 (Cox 2 )蛋白表达 ,采用四唑氮蓝 (MTT)比色法检测细胞增殖 ,流式细胞仪测定细胞周期和凋亡 ,酶联免疫吸附试验 (ELISA)检测前列腺素E2 (PGE2 )含量。结果 Celecoxib可下调GBC SD细胞Cox 2蛋白表达。Celecoxib抑制GBC SD细胞增殖作用呈时间依赖性 ,作用 2天后就有轻度抑制 ,第 3、第 4和第 5天抑制作用已相当明显 (P <0 .0 5 ,与对照组相比 ) ,并且这种增殖抑制作用能被 2 0 0 pg/mlPGE2 拮抗。Celecoxib诱导GBC SD细胞G1 S期阻滞 ,4 0 μmol/L(P <0 .0 5 )和2 0 μmol/L(P <0 .0 5 )Celecoxib组G0 G1期细胞含量比对照组 ( 0 μmol/L)明显增高 ,而S期和G2 /M期细胞含量比对照组明显降低 (P <0 .0 5 )。 4 0 μmol/L(P <0 .0 5 )和 2 0 μmol/L(P <0 .0 5 )Celecoxib处理 4 8h后GBC SD细胞凋亡率明显上升。 结论 Celecoxib通过Cox 2和PGE2 途径抑制GBC SD细胞生长和诱导凋亡。

Objective To evaluate the effects and mechanisms of celecoxib in inducing proliferation inhibition and apoptosis on human gallbladder cancer cells GBC SD. Methods Cox 2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The anti proliferative effect was measured by methabenzthiazuron (MTT) assay. Cell cycle and apoptosis were analyzed by flow cytometry. The PGE 2 levels in the supernatant of cultured GBC SD cells were quantified by enzyme linked immunoabsordent assay (ELISA). Results Celecoxib down regulated the Cox 2 protein expression of GBC SD cells. Celecoxib suppressed the production of PGE 2 and inhibited growth of GBC SD cells; and the anti proliferative effect of celecoxib could be abolished by addition of 200 pg/mL PGE 2 . Celecoxib induced proliferation inhibition and apoptosis by G 1 S cell cycle arrest. Conclusion Cyclooxygenase 2 specific inhibitor celecoxib inhibits proliferation and induces apoptosis of human carcinoma of gallbladder cells via suppression of PGE 2 production in vitro.