蓝萼乙素通过NF-κB信号通路调控舌鳞状细胞癌细胞的增殖与凋亡

Glaucocalyxin B Regulating the Proliferation and Apoptosis of Tongue Squamous Cell Carcinoma Cells through NF-κB Signaling Pathway

目的:探讨蓝萼乙素调控舌鳞状细胞癌(TSCC)细胞增殖、凋亡的分子机制。方法:体外培养人TSCC细胞SCC15,使用不同浓度的(3,6,12μmol·L-1)蓝萼乙素处理24 h。甲基噻唑基四唑(MTT)检测细胞增殖;流式细胞术检测细胞凋亡率;蛋白免疫印迹法(Western blot)检测蓝萼乙素对核因子-κB(NF-κB)信号通路相关蛋白[磷酸化核因子κB抑制蛋白α(p-IκBα)、磷酸化p65(p-p65)、p65、核因子κB抑制蛋白α(IκBα)]表达的影响;NF-κB信号通路的激活剂脂多糖(LPS)预处理SCC15细胞后,MTT法与流式细胞术分别检测细胞增殖能力及细胞凋亡率;Western blot法检测细胞周期蛋白1(CyclinD1)、增殖细胞核抗原(PCNA)、B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、半胱氨酰天冬氨酸特异性蛋白酶3总蛋白(T-caspase-3)的蛋白水平。结果:与空白组相比,蓝萼乙素不同浓度组细胞增殖率显著降低,CyclinD1、PCNA、Bcl-2、p-IκBα、p-p65蛋白水平显著降低,细胞凋亡率显著升高,Bax、Cleaved caspase-3蛋白水平显著升高(P<0.05);与蓝萼乙素高浓度+PBS组相比,蓝萼乙素高浓度+LPS组细胞增殖率显著升高,CyclinD1、PCNA、Bcl-2蛋白水平显著升高,细胞凋亡率显著降低,Bax、Cleaved caspase-3蛋白水平显著降低(P<0.05)。结论:蓝萼乙素能够抑制TSCC细胞增殖,促进细胞凋亡,其机制可能是通过抑制NF-κB信号通路的活化而发挥作用。

Objective:To explore the molecular mechanism of glaucocalyxin B in regulating the proliferation and apoptosis of TSCC cells.Methods:Human TSCC cells SCC15 were cultured in vitro and treated with different concentrations(3,6 and 12μmol·L-1)of glaucocalyxin B for 24 h.MTT was used to detect the cell proliferation.Flow cytometry was used to detect the apoptosis rate.Western blot was used to detect the effect of glaucocalyxin B on the expressions of NF-κB signaling pathway related proteins p-IκBα,p-p65,p65 and IκBα.After the pretreatment of SCC15 cells with LPS,an activator of NF-κB signaling pathway,MTT assay and flow cytometry was used to detect the cell proliferation and the apoptosis rate,respectively.Western blot was used to detect the protein levels of CyclinD1,PCNA,Bcl-2,Bax,Cleaved Caspase-3 and T-caspase-3.Results:Compared with that of the blank group,the cell proliferation rate of glaucocalyxin B groups at different concentrations was significantly reduced,the protein levels of CyclinD1,PCNA,Bcl-2,p-IκBαand p-p65 were significantly reduced,the apoptosis rate was significantly increased,and the protein levels of Bax and Cleaved caspase-3 were significantly increased(P<0.05).Compared with that of glaucocalyxin B high concentration+PBS group,the cell proliferation rate of glaucocalyxin B high concentration+LPS group was significantly increased,and the protein levels of CyclinD1,PCNA,and Bcl-2 were significantly increased,while the apoptosis rate was significantly reduced and the protein levels of Bax and Cleaved caspase-3 were significantly reduced(P<0.05).Conclusion:Glaucocalyxin B can inhibit the proliferation of TSCC cells and promote the apoptosis.The mechanism may be through inhibiting the activation of NF-κB signaling pathway.