摘要
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 a selective inhibitor of COX-2 nimesulide on growth and apoptosis of human cholangiocarcinoma cell line QBC939 in vitro. Methods MTT assay was used to determine the influence of nimesulide on the proliferation of QBC939 cells, apoptosis of QBC939 was measured by transmission electron microscopy and flow cytometry. Expression of apeptosis related genes mRNA and bcl-2,bax, survivin were detected by RT-PCR and immunocytochemistry. Results Nimesulide effects a dose-dependent and time-dependent growth inhibition on QBC939 cells. High concentration of nimesulide （200μmol/L） not only inhibits the growth of QBC939 cells but also induces apoptosis cell nuclear condensation and apeptotic bodies were seen by transmission electron microscopy. Immunocytochemistry and RT-PCR shows upregulation of bax and down regulation of bcl-2 and survivin. Conclusion Nimesulide significantly inhibits the proliferation of QBC939 in vitro by induction of apeptosis in a dose- and time- dependent manner.
出处
《中华普通外科杂志》
CSCD
北大核心
2006年第3期203-205,共3页
Chinese Journal of General Surgery
关键词
胆管肿瘤
药物筛选试验
抗肿瘤
动物实验替代试验
脂氧合酶抑制剂
Bile duct neoplasms
Drug screening assays, antitumor
Animal testing alternatives
Lipoxygenase inhibitors
