白花丹素通过ROS介导的JNK通路诱导舌鳞状细胞癌细胞凋亡

Plumbagin Induces Tongue Squamous Cell Carcinoma Cells Apoptosis via ROS-mediated JNK Signaling Pathway

目的:探究白花丹素诱导舌鳞状细胞癌细胞凋亡的分子机制。方法:CCK-8实验检测白花丹素对舌鳞状细胞癌细胞增殖的影响,流式细胞术检测舌鳞状细胞癌细胞凋亡率和细胞内活性氧水平,Western blot检测Bcl-2、Bax和p-JNK蛋白的表达。结果:CCK-8显示白花丹素有显著的抗舌鳞状细胞癌细胞增殖作用(P<0.001)。流式细胞术显示白花丹素可呈剂量依赖性地促进舌鳞状细胞癌细胞凋亡(P<0.001),有效刺激舌鳞状细胞癌细胞内产生活性氧(P<0.001)。Western blot结果显示,白花丹素作用后Bax/Bcl-2的比值升高(P<0.01,P<0.001),p-JNK蛋白表达随着作用时间延长逐渐升高(P<0.05,P<0.01,P<0.001)。然而,当我们使用活性氧清除剂NAC后,p-JNK蛋白表达和细胞凋亡率都被显著逆转(P<0.001,P<0.001)。结论:白花丹素可以有效抑制舌鳞状细胞癌细胞的增殖,此外白花丹素可以通过活性氧介导的JNK途径诱导舌鳞状细胞癌细胞凋亡。

Objective:To explore the underlying molecular mechanisms of plumbagin-induced apoptosis of tongue squamous cell carcinoma cells.Methods:The effect of plumbagin on cell proliferation was determined by CCK-8 assay.Cell apoptosis and the expression of cellular ROS generation were measured by flow cytometry.Protein levels of Bcl-2,Bax,and p-JNK were evaluated by Western blot.Results:CCK-8 assay showed that plumbagin significantly inhibited cell proliferation in a dose-dependent manner(P<0.001).The results of flow cytometer suggested that plumbagin markedly increased cell apoptosis and ROS generation.Western blot showed that the expression of Bax increased,while the expression of Bcl-2 decreased after plumbagin treatment.In a time-dependent manner,plumbagin up-regulated the expressions of p-JNK(P<0.05,P<0.01,P<0.001).However,the ROS inhibitor NAC significantly reversed the expression level of p-JNK and cell apoptosis(P<0.001,P<0.001).Conclusion:Plumbagin had excellent anti-proliferation effects on tongue squamous cell carcinoma cells.In addition,plumbagin induced cell apoptosis through ROS-mediated JNK pathway.