丹参酮ⅡA免疫纳米粒调控Wnt/β-catenin信号通路抑制肝癌干细胞侵袭转移的作用机制

Mechanisms of Tanshinone ⅡA immune nanoparticles on inhibiting invasion and metastasis of liver cancer stem cells through Wnt/β-catenin signaling pathway

目的探究丹参酮ⅡA免疫纳米粒(TSIIA-mPEG-PLGA-PLL-EpCAM NPs)通过Wnt/β-catenin信号通路抑制肝癌干细胞侵袭转移的作用机制。方法构建丹参酮ⅡA免疫纳米粒。划痕实验与Transwell实验分别检测丹参酮ⅡA免疫纳米粒对肝癌干细胞侵袭转移作用的影响。采用Western blot分析丹参酮ⅡA免疫纳米粒作用肝癌干细胞后,β-catenin及其破坏复合体GSK3β、Axin、APC的蛋白表达水平;荧光定量PCR检测上游信号因子Wnt1 mRNA表达水平。结果与丹参酮ⅡA相比,丹参酮ⅡA免疫纳米粒抑制肝癌干细胞的侵袭转移效果更佳(P<0.01)。丹参酮ⅡA免疫纳米粒能够通过抑制Wnt1的表达(P<0.01),抑制β-catenin的入核(P<0.01),同时降低GSK3β、Axin、APC的蛋白表达水平(P<0.01)。结论丹参酮ⅡA免疫纳米粒可能通过抑制Wnt/β-catenin信号通路,降低GSK3β、Axin、APC蛋白的表达,从而抑制肝癌干细胞的侵袭转移。

Objective To explore the mechanism of TanshinoneⅡA immune nanoparticles(TSⅡA-mPEG-PLGA-PLL-EpCAM NPs)on inhibiting invasion and metastasis of liver cancer stem cells through Wnt/β-catenin signaling pathway.Methods TanshinoneⅡA immune nanoparticles were constructed.The effects of TanshinoneⅡA immune nanoparticles on the invasion and metastasis of liver cancer stem cells were detected by Scratch test and Transwell test.After TanshinoneⅡA immune nanoparticles acted on liver cancer stem cells,the protein expressions ofβ-catenin and its destruction complex GSK3β,Axin and APC were analyzed by Western blot.The mRNA expression of upstream signal factor Wnt1 was detected by fluorescence quantitative PCR.Results Compared with TanshinoneⅡA,the effect of TanshinoneⅡA immune nanoparticles on inhibiting the invasion and metastasis of liver cancer stem cells was more significant(P<0.01).TanshinoneⅡA immune nanoparticles could inhibit the expression of Wnt1(P<0.01)to reduce the entry ofβ-catenin into the nucleus(P<0.01),and decrease the protein expressions of GSK3β,Axin and APC(P<0.01).Conclusion TanshinoneⅡA immune nanoparticles may inhibit the invasion and metastasis of liver cancer stem cells by decreasing the protein expressions of GSK3β,Axin and APC via Wnt/β-catenin signaling pathway.