亚砷酸钠对PC12细胞增殖、凋亡及Hippo-YAP通路的影响

Effects of sodium arsenite on proliferation,apoptosis and Hippo-YAP pathway of PC12 cells

目的探讨亚砷酸钠(NaAsO_(2))对PC12细胞增殖、凋亡及Hippo-YAP信号通路的影响。方法以PC12细胞为研究对象,分别设立对照组、不同浓度(20、40、60μmol·L^(-1))NaAsO_(2)处理组。流式细胞术、Western blot法来检测NaAsO_(2)对PC12细胞周期、细胞凋亡率、细胞凋亡相关蛋白(p53、BAX、BCL-2)以及Hippo-YAP信号通路相关蛋白(MST1、p-MST1、LATS1、p-LATS1、YAP和p-YAP)的影响。结果与对照组相比,40μmol·L^(-1) NaAsO_(2)处理24 h后,PC12细胞中G_(0)/G_(1)期比例明显降低(P<0.01),而S期比例明显增加(P<0.05),60μmol·L^(-1) NaAsO_(2)明显降低G_(0)/G_(1)期、G_(2)/M期比例(P<0.05),而增加S期比例(P<0.01);细胞凋亡率及凋亡相关蛋白BAX和p53的表达被40μmol·L^(-1) NaAsO_(2)明显增加(P<0.05),而BCL-2蛋白表达被明显降低(P<0.01);60μmol·L^(-1) NaAsO_(2)明显降低MST1、LATS1、YAP蛋白表达(P<0.05);不同浓度的NaAsO_(2)则明显上调p-MST1、p-LATS1、p-YAP蛋白表达(P<0.05)。结论NaAsO_(2)可干扰PC12细胞周期、诱导PC12细胞凋亡、激活Hippo-YAP信号通路。

Aim To investigate the effects of sodium arsenite(NaAsO_(2))on the proliferation,apoptosis and Hippo-YAP signaling pathway of PC12 cells.Methods PC12 cells were randomly divided into four groups:the control group and the NaAsO_(2) groups(incubated with 20,40,and 60μmol·L^(-1) NaAsO_(2),respectively).Flow cytometry and Western blotting were used to detect the effects of NaAsO_(2) on the cell cycle,apoptotic rates,apoptosis related proteins(p53,BAX,BCL-2),and Hippo-YAP signaling pathway-related proteins(MST1,p-MST1,LATS1,p-LATS1,YAP,p-YAP).Results Compared with control group,after 40μmol·L^(-1) NaAsO_(2) treatment for 24 h,the G_(0)/G_(1) phase ratio in PC12 cells significantly decreased(P<0.01),while the S phase ratio significantly increased(P<0.05)while 60μmol·L^(-1) NaAsO_(2) significantly decreased the ratio of G_(0)/G_(1) phase and G_(2)/M phase(P<0.05),and increased the ratio of S phase(P<0.01).The apoptotic rate and the expression level of apoptosis-related protein BAX and p53 were up-regulated significantly(P<0.05),but the expression of BCL-2 protein was significantly down-regulated by 40μmol·L^(-1) NaAsO_(2)(P<0.01).Meanwhile,60μmol·L^(-1) NaAsO_(2) significantly decreased the protein expressions of MST1,LATS1,and YAP(P<0.05).However,the protein expressions of p-MST1,p-LATS1,and p-YAP were significantly up-regulated by different concentrations of NaAsO_(2)(P<0.05).Conclusions NaAsO_(2) can interfere with the cell cycle,induce the cell apoptosis,and activate the Hippo-YAP signaling pathway in PC12 cells.