【摘要】 目的探讨缺氧对舌苔形成相关细胞凋亡的影响。方法将Co Cl2作用于撤血清舌鳞癌Tca-8113细胞,建立化学性缺氧模型。MTT法检测细胞增殖活性,荧光染色检测细胞凋亡,琼脂糖电泳观察DNA Ladder,流式细胞术检测细胞周期分布和凋亡,Western blotting检测Akt、NF-κB p50、NF-κB p65、Bax、Bcl-2、HSP 70的表达水平。结果 50~400μmol/L Co Cl2作用细胞8~24h能明显抑制撤血清舌鳞癌细胞增殖(P<0.05),并具有量效和时效关系。Hoechst荧光染色显示Co Cl2可引起撤血清舌鳞癌细胞凋亡。流式细胞术显示100,200,400μmol/L Co Cl212h缺氧-12h复氧可促进撤血清舌鳞癌细胞凋亡(P<0.05),且凋亡率随Co Cl2的浓度增高。Western blotting表明12h缺氧-12h复氧可上调Bax、HSP 70、NF-κB p50,下调NF-κB P65表达(100μmol/L Co Cl2),促进舌鳞癌细胞凋亡。结论缺氧是促进舌苔形成相关细胞凋亡的重要因素,其发生机制可能受NF-κB、Bax、HSP 70蛋白的调控。更多还原

【Abstract】 Objective To investigate the hypoxia- induced apoptosis in tongue coating- related cells. Methods Cobalt chloride( Co Cl2),a chemical hypoxia inducer,was added in Tca- 8113 cells with serum withdrawal to develop a hypoxic model. Proliferative activity of Tca- 8113 cell was detected by MTT assay. Apoptosis were detected by Hoechst 33342 staining. DNA fragments in Tca- 8113 cells were analyzed by gel electrophoresis for visualizations of apoptosis- indicative DNA ladders. The distribution of cell cycle and apoptosis were detected by flow cytometric analysis. The expressions of Akt,natural factor- κB( NF-κB) p50,NF- κB p65,Bax,Bcl- 2 and heat shock proteins( HSP) 70 were detected by Western blot. Results Compared with control group,proliferative activity of Tca- 8113 cells decreased after Co Cl2( 50 ~ 400μmol/L) acting for 8 hours( P < 0. 05) in a dose- effect dependent and time- effect dependent manner. Hoechst 33342 staining showed that Co Cl2 promoted apoptotic rate of Tca- 8113 cells. Treated with Co Cl2( 100,200,400μmol/L) in 12 h hypoxia- 12 h reoxygenation,apoptotic rate of Tca-8113 cells increased significantly( P < 0. 05) in a dose- effect dependent manner. Western blot analysis showed that 12 h hypoxia- 12 h reoxygenation lead to increase of Bax,HSP70 and NF- κB p50( 100μmol / L Co Cl2) but not Akt and Bcl- 2,and decrease of NF- κB p65( 100μmol / L Co Cl2). Conclusion Hypoxia is an important factor in the apoptosis of tongue coating- related cells. And its mechanism may through regulation of Bax,HSP 70 and NF- κB.更多还原