环氧化酶-2 抑制剂Nimesulide对膀胱癌T24细胞体外生长的抑制作用

 目的 观察环氧化酶-2（COX-2）抑制剂Nimesulide（NIM）对人膀胱癌T24细胞株体外生长的影响，并探讨其作用机制。方法 采用细胞形态学、MTT法、流式细胞术、吖啶橙-溴化乙啶双染法（AO-EB）等技术检测NIM对体外培养的人膀胱癌细胞株T24生长的抑制效应。结果 细胞形态学观察可见NIM对T24细胞生长具有明显的细胞毒作用；MTT法、流式细胞术分析显示NIM能明显抑制T24细胞的增殖，且呈剂量、时间依赖关系；细胞周期分析表明，NIM能使T24细胞G0／G1期细胞比例升高，S期和G2／M期细胞比例下降；流式细胞DNA直方图上出现典型的亚二倍体“凋亡峰”，细胞凋亡率与NIM作用时间和剂量相关；荧光显微镜检查显示NIM处理的T24细胞AO-EB双染色后，细胞核内可见浓染致密的颗粒荧光，典型细胞可见新月形改变，固缩或片段化的核。结论 COX-2抑制剂NIM对T24细胞生长具有明显的抑制作用，其作用机制可能与阻断细胞周期、诱导细胞凋亡等有关。

The Inhibit Effect of the Cyclooxygenase-2 Inhibitor Nimesulide on Human Bladder Cancer Cell Line T24 in Vitro

Objective To investigate the growth effects of the selective cyclooxygenase-2 inhibitor Nimesulide on human bladder cancer cell line T24 in vitro, and to clarify its probable mechanism. Methods The T24 cell line was culture in vitro, and exposed to a range of concentrations of Nimesulide at different time. The effect of COX-2 inhibitor Nimesulide on the proliferation and apoptosis of human bladder cancer cell line T24 in vitro were observed by using cell morphology , MTF assay , flow cytometry, fluorescent microscopy. Results Significant cytotoxicity of cyclooxygenase-2 inhibitor Nimesulide on human bladder cancer cell line T24 were observed by cell morphology. MTT assay and flow cytometry show that cyclooxygenase-2 inhibitor Nimesulide significantly inhibited cell proliferation of T24 cell line in vitro in a dose- and time-dependent manner, Cell-cycle analysis by flow cytometry suggested that Nimesulide induced a decrease in S and G2/M phase population and a significant increase in G0/G1 population compared with untreated T24 cells. And a typical subdiploid peak before Phase G0/G1 was detected by flow cytometry. The rates of apoptosis were correlated with the concentration of Nimesulide and time of action. A typical apoptosis morphology were also observed by fluorescent microscopy. Conclusion The selective cyclooxygenase-2 inhibitor Nimesulide has significant inhibit effects on the growth of T24 cell, which is closely related to block cell cycle and apoptosis-inducing.