选择性抑制剂celecoxib诱导肝癌细胞凋亡及其相关分子机制的实验研究

Experimental Studies on Celecoxib-induced Hepatoma Cell Apoptosis and Its Molecular Mechanism

目的:研究环氧合酶-2(COX-2)选择性抑制剂celecoxib抑制肝癌细胞增殖和诱导肝癌细胞凋亡过程中的作用及其可能的机制。方法:采用celecoxib作用于Bel-7402、SMMC-7721的2种肝癌细胞株,以四甲基偶氮唑蓝(MTT)法测定肿瘤细胞的增殖抑制率、TUNEL实验、流式细胞术检测细胞凋亡率、免疫细胞化学和Westetm blot实验测定细胞凋亡及Akt信号转导途径相关蛋白的激活状态。结果:MTT法显示celecoxib可抑制上述两株肝癌细胞增殖;TUNEL实验及流式细胞术结果证实,celecoxib处理后的肿瘤细胞可发生细胞凋亡;免疫细胞化学及Western blot实验显示,celecoxih诱导凋亡主要是通过抑制Akt的Thr308磷酸化,进而激活caspase-9、caspase-3进行的。结论:celecoxib明显抑制细胞增殖、诱导细胞凋亡,其主要机制可能通过Akt信号转导途径、激活caspase通路实现的。

To investigate the cell growth inhibition and apoptosis induced by selective Cyclooxyygenare-2(COX-2) inhibitor celecoxib and its molecular mechanisms. Methods: Two human hepatocarcinoma cell strains, Bel-7402 and SMMC-7721, were cultured and treated with celecoxib. Cell growth rates were assessed by monotetrazdium (MTT) colorimetric assay. Apoptosis was examined by TUNEL and flow cytometry. Immunohistochemistry with imaging analysis was used to determine COX-2 expression and Akt phosphorylation level. The expressions of caspase-3, caspase-9 and Akt phosphorylation were also examined by Western Blot analysis. Results: The celecoxib inhibited the proliferation and induced apoptosis of the two hepatocarcinoma cell strains in a dose and time dependent manner. Remarkable activation of caspase and significant reduction of Akt (Thr308) phosphorylation could be observed by Western blotting. Conclusion: The celecoxib inhibited the proliferation and induced apoptosis of the Bel-7402 and SMMC-7721 cells in a dose and time dependent manner and induced cell apoptosis by the mechanism of Akt/caspase pathway - dependent signal transduction.