克班宁通过DUSP5/ERK信号通路抑制胰腺癌细胞生长

Crebanine inhibits pancreatic cancer cell growth through DUSP5/ERK signaling pathway

目的:研究克班宁对胰腺癌细胞增殖和凋亡的影响,并探讨其潜在的作用机制。方法:采用CCK8、克隆形成实验和EdU染色检测克班宁对人胰腺癌细胞Panc-1和Patu-8988增殖的影响;采用流式细胞术和TUNEL染色检测克班宁对细胞凋亡的影响;采用Western blot实验检测克班宁对DUSP5蛋白,增殖相关蛋白Ki67,凋亡相关蛋白Bcl-2和Bax,以及p-ERK1/2蛋白的影响。结果:克班宁处理24 h后的胰腺癌细胞增殖能力显著降低(P<0.05),而凋亡率明显升高(P<0.05);同时,DUSP5的表达上调(P<0.05),p-ERK1/2的表达下降(P<0.05)。敲低DUSP5能够有效逆转克班宁对胰腺癌细胞的抑增殖和促凋亡作用(均P<0.05),还能部分恢复克班宁降低的p-ERK1/2的蛋白水平(P<0.05)。结论:克班宁能够抑制胰腺癌细胞增殖并促进细胞凋亡,其机制是促进DUSP5的表达进而抑制ERK1/2的激活。

Objective:To explore the effect of crebanine on the proliferation and apoptosis of pancreatic cancer cells and its potential mechanism.Methods:CCK8,clone formation assay and EdU staining were used to detect the effect of crebanine on the proliferation of human pancreatic cancer cells Panc-1 and Patu-8988.The effect of crebanine on apoptosis was detected by flow cytometry and TUNEL staining.Western blot assay was conducted to determine the effects of crebanine on DUSP5 protein,proliferation-related protein Ki67,apoptosis-related proteins Bcl-2 and Bax,and p-ERK1/2.Results:After 24 h treatment with crebanine,the proliferation ability of pancreatic cancer cells decreased(P<0.05),while the apoptosis rate increased significantly(P<0.05).Meanwhile,the expression of DUSP5 was markedly upregulated(P<0.05),and the expression of p-ERK1/2 was downregulated(P<0.05).Knockdown of DUSP5 effectively reversed the anti-proliferation and pro-apoptotic abilities of crebanine in pancreatic cancer cells(all P<0.05),and partially restored the p-ERK1/2 protein reduced by crebanine(P<0.05).Conclusion:Crebanine inhibits the proliferation and promotes apoptosis of pancreatic cancer cells by upregulating DUSP5 expression and subsequently inactivating ERK1/2 signaling.