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目的研究选择性环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂尼美舒利对人胆管癌细胞系QBC939增殖和凋亡的影响。方法应用MTT比色法观察尼美舒利对人胆管癌细胞系 QBC939增殖的影响,细胞凋亡采用透射电镜及流式细胞仪检测,应用免疫组化、RT-PCR检测尼美舒利对凋亡相关基因表达的影响。结果尼美舒利呈时间、剂量依赖性抑制胆管癌细胞增殖,高浓度 (200μmol／L)尼美舒利不仅抑制胆管癌细胞增殖,而且诱导其凋亡,流式细胞仪研究显示,随药物浓度增加,细胞凋亡率显著增加,透射电镜下可见典型的细胞凋亡形态学改变:细胞核染色质致密浓缩, 凋亡小体形成等,免疫组化和RT-PCR测定显示尼美舒利显著下调bcl-2、survivin表达,而bax表达上调。结论尼美舒利显著抑制QBC939细胞增殖,诱导其凋亡,且呈时间、剂量依赖性,bcl-2、bax、 survivin等凋亡相关基因可能参与了尼美舒利诱导的QBC939细胞凋亡过程。
Objective To investigate the effect of selective cyclooxygenase-2 (COX-2) inhibitor nimesulide on proliferation and apoptosis of human cholangiocarcinoma cell line QBC939. Methods The effect of nimesulide on proliferation of human cholangiocarcinoma cell line QBC939 was observed by MTT assay. Apoptosis was detected by transmission electron microscope and flow cytometry. Nimesulide was detected by immunohistochemistry and RT-PCR. The effects of expression of death-related genes. Results Nimesulide inhibited the proliferation of cholangiocarcinoma cells in a dose- and time-dependent manner. Nimesulide at a high concentration (200μmol/L) not only inhibited the proliferation of cholangiocarcinoma cells, but also induced its apoptosis. Flow cytometry studies showed that with the drug The increase in the concentration of apoptotic cells increased significantly. Typical morphology of apoptosis was observed under transmission electron microscope: dense nuclear chromatin and apoptotic bodies were formed. Immunohistochemistry and RT-PCR showed that nimesulide was significant. Down-regulation of bcl-2 and survivin expression, while bax expression was up-regulated. Conclusion Nimesulide significantly inhibits the proliferation and induces apoptosis of QBC939 cells in a dose and time-dependent manner. Apoptosis-related genes such as bcl-2, bax and survivin may be involved in the apoptosis of QBC939 cells induced by nimesulide. .