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目的研究环加氧酶-2(cyclooxygenase-2,COX-2)抑制剂NS398抑制人骨肉瘤细胞MG-63增殖及诱导其凋亡的效应和机制。方法MTT比色法观察NS398的细胞毒性作用及其浓度依赖性;透射电子显微镜观察细胞凋亡的形态学变化;琼脂糖凝胶电泳观察凋亡细胞DNA Ladder形成。流式细胞仪检测细胞凋亡率及与NS398作用时间的关系;Western blot测定bcl-2蛋白表达。结果不同浓度(0.1~100μmol/L)NS398均可使细胞生长受到抑制;电镜显示细胞呈典型的凋亡形态学变化;琼脂糖凝胶电泳显示凋亡细胞DNA Ladder形成;流式细胞术检测证实细胞凋亡率与NS398作用时间呈明显相关。Western blot检测显示随着NS398作用时间延长,可不同程度地降低bcl-2蛋白表达。结论NS398能抑制人MG-63细胞增殖,诱导细胞凋亡,其作用机制可能与调控bcl-2蛋白表达有关。
Objective To study the effect of cyclooxygenase-2 (COX-2) inhibitor NS398 on the proliferation and apoptosis of human osteosarcoma MG-63 cells. Methods The cytotoxicity and concentration dependence of NS398 were observed by MTT assay. Morphological changes of apoptotic cells were observed by transmission electron microscopy. The DNA ladder was observed by agarose gel electrophoresis. Flow cytometry was used to detect the apoptosis rate and the relationship with NS398 action time. Western blot was used to detect the expression of bcl-2 protein. Results The cell growth was inhibited by NS398 at different concentrations (0.1-100 μmol / L). The morphological changes of cells were observed by electron microscopy. The DNA ladder of apoptotic cells was observed by agarose gel electrophoresis. Flow cytometry confirmed Apoptosis rate and NS398 role of time was significantly correlated. Western blot showed that with the prolongation of NS398, the expression of bcl-2 protein could be decreased to some extent. Conclusion NS398 can inhibit the proliferation and induce the apoptosis of human MG-63 cells. Its mechanism may be related to the regulation of bcl-2 protein expression.