一氧化氮诱导口腔鳞癌细胞凋亡和 P53蛋白表达的实验研究

目的：研究外源性一氧化氮（NO）对人舌鳞癌Tca8113细胞诱导凋亡的作用及可能机制。方法：硝普钠（SNP）作为NO的供体，用不同时间和不同浓度SNP处理细胞。采用MTT法检测N（）对细胞的生长抑制，丫啶橙染色、Wright—Giemsa染色、流式细胞仪、琼脂糖凝胶电泳观察细胞凋亡作用，蛋白电泳法分析细胞凋亡机制。结果：NO对Tca8113细胞的抑制作用呈剂量和时间依赖性。SNP处理细胞后，DNA、RNA合成减弱；细胞形态学出现凋亡的特征性改变；电泳可见典型的DNA Ladder形成；流式细胞仪上见随着SNP浓度增高，细胞凋亡随之增加，并出现G2／M期阻滞；Western blot显示随着SNP浓度增高，P53蛋白表达水平上调。结论：外源性NO对人舌鳞癌Tca8113细胞有增殖抑制和诱导凋亡作用，P53蛋白介入了硝普钠引起的细胞凋亡调控作用。

Apoptosis induces by exogenous nitric oxide in Tca8113 cells

Objective: To observe the apoptosis induced by exogenous NO in Tca8113 cells and to investigate the possible mechanism.Methods: SNP as NO donor was used to treat the tongue squamous cell carcinoma Tca8113 cells.Cytotoxic and apoptotic effects of NO on Tca8113 cells were examined by using MTT assay,acridine orange(AO) staining,Wright-Giemsa staining,agarose gel electrophoresis and flow cytometry.Western blot was performed for investigating the apoptotic mechanism.Results: NO had a remarkable proliferation inhibiting effect on Tca8113 cells.After being exposed to exogenous NO,Tca8113 cells showed series of apoptotic morphological changes such as cell shrinkage,nuclear condensation;and also showed DNA fragmentation,G2/M phase arrest as well as upregulation of the tumor suppressor P53 protein.Conclusion: Exogenous NO has a proliferation inhibition and apoptosis induction effect on Tca8113 cells in a concentration and time-dependent manner,P53 protein may be involved in the apoptosis induced by NO.