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目的:探讨NF-κB信号转导通路在紫草素诱导Tca-8113细胞凋亡中的作用。方法:采用蛋白印迹法检测IκBa,磷酸化-IκBa、bcl-2及Bax蛋白的表达,采用EMSA方法检测NF-κB的DNA结合活性,采用酶联免疫吸附方法(ELISA)分析Caspase 3、8、9的活性。采用SPSS12.0软件包对数据进行单因素方差分析及t检验。结果:经过紫草素作用的癌细胞,磷酸化-IκBa蛋白及NF-κB的DNA结合活性均明显降低,Bcl-2蛋白表达显著减少。Caspase 3、8、9在紫草素诱导的细胞凋亡过程中被激活,泛Caspase阻断剂Z-Asp-CH2-DCB可以明显抑制紫草素引起的细胞凋亡(P=0.02)。结论:紫草素诱导口腔鳞癌细胞的凋亡作用,至少部分通过抑制NF-κB信号通路活性,继而调节其下游调亡调控分子,包括bcl-2家族及Caspase家族等来实现。应用紫草素特异性抑制口腔鳞癌中高激活状态的NF-κB通路,可望成为口腔鳞癌防治的一条新的有效途径。
Objective: To investigate the role of NF-κB signal transduction pathway in shikonin-induced Tca-8113 cell apoptosis. Methods: The expression of IκBa, phospho-IκBa, bcl-2 and Bax proteins were detected by Western blotting. The DNA binding activity of NF-κB was detected by EMSA. The expressions of Caspase 3, 9 activity. SPSS12.0 software package was used to analyze the data by one-way ANOVA and t-test. Results: DNA binding activity of phosphorylated-IκBa protein and NF-κB decreased significantly and the expression of Bcl-2 protein decreased significantly after transfected with shikonin. Caspase 3, 8 and 9 were activated during shikonin-induced apoptosis, and pan-Caspase inhibitor Z-Asp-CH2-DCB significantly inhibited shikonin-induced apoptosis (P = 0.02). CONCLUSION: Shikonin can induce the apoptosis of oral squamous cell carcinoma cells, at least partially by inhibiting the activity of NF-κB signaling pathway and regulating its downstream apoptosis regulatory molecules, including bcl-2 family and Caspase family. The application of shikonin to specifically inhibit NF-κB pathway in oral squamous cell carcinoma is expected to be a new effective approach for the prevention and treatment of oral squamous cell carcinoma.