TLR4-NFκB信号通路在顺铂致胃癌SGC-7901细胞凋亡中的作用

The role of TLR4-NFκB signaling pathway in cisplatin-induced gastric cancer SGC-7901 cells apoptosis

目的探讨TLR4-NFκB信号通路在顺铂致胃癌细胞凋亡中的作用。方法终浓度为10μmol/L的顺铂处理SGC-7901细胞12 h后,Western blot检测SGC-7901细胞中TLR4和NFκB的表达情况。设计合成无义序列和TLR4 siRNA序列,分别转染至SGC-7901细胞,即无义序列转染组(Scramb-Anti-TLR4 group)和TLR4 siRNA序列转染组(Anti-TLR4 group)。顺铂处理后,Western blot检测细胞中TLR4、NFκB、Bax、Bcl-2、Activecaspase-3和GAPDH的表达,流式细胞术检测细胞凋亡和坏死。结果顺铂处理后TLR4(t=14. 070,P <0. 001)和NFκB(t=17. 660,P <0. 001)的表达显著增加。与Scramb-Anti-TLR4组相比较,Anti-TLR4组在顺铂处理后TLR4(t=16. 810,P <0. 001)和NFκB(t=15. 040,P <0. 001)表达显著下降; Bax(t=17. 870,P <0. 001)和Activecaspase-3(t=8. 881,P <0. 001)表达显著升高,Bcl-2(t=17. 300,P <0. 001)表达显著下降;细胞凋亡率(t=4. 122,P <0. 001)和坏死率(t=9. 367,P <0. 001)显著升高。结论顺铂可以激活胃癌SGC-7901细胞的TLR4-NFκB信号通路,抑制TLR4-NFκB信号通路可以增强顺铂的致凋亡效应。

Objective To explore the role of TLR4-NFκB signaling pathway in cisplatin-induced gastric cancer SGC-7901 cells apoptosis. Methods SGC-7901 cells were treated with cisplatin at a final concentration of 10μmol/L for 12 h. Western blot was used to detect the expression of TLR4 and NFκB in SGC-7901 cells. Nonsense sequences and TLR4 siRNA sequences were designed,synthesized and transfected into SGC-7901 cells,named Scramb-Anti-TLR4 group and Anti-TLR4 group. After the cells were treated with cisplatin,the expression of TLR4,NFκB,Bax,Bcl-2,Active caspase-3 and GAPDH were detected by Western blot. Apoptosis and necrosis were detected by flow cytometry. Results The expression of TLR4( t = 14. 070,P < 0. 001) and NFκB( t = 17.660,P < 0. 001) were significantly increased after cisplatin treatment. Compared with the Scramb-Anti-TLR4 group,the expression of TLR4( t = 16. 810,P < 0. 001) and NFκB( t = 15. 040,P < 0. 001) were significantly decreased in the Anti-TLR4 group after cisplatin treatment. The expression of Bax( t = 17. 870,P < 0. 001) and Active caspase-3( t = 8. 881,P < 0. 001) were significantly increased. The expression of Bcl-2( t = 17. 300,P < 0.001) was significantly decreased. Apoptosis rate( t = 4. 122,P < 0. 001) and necrosis rates( t = 9. 367,P < 0. 001)were significantly increased. Conclusion Cisplatin can activate TLR4-NFκB signaling pathway in gastric cancer SGC-7901 cells. Inhibition of TLR4-NFκB signaling pathway can enhance cisplatin-induced apoptotic effect.