红景天苷对肾癌细胞A498侵袭、免疫炎症因子及ERK1/2和STAT3活化的影响

Effect of salidroside on A498 invasion,immune inflammatory cytokines and activation of ERK1/2 and STAT3 in renal carcinoma cells

目的:探索红景天苷(SAL)对肾癌细胞A498侵袭、免疫炎症因子及细胞外信号调节激酶1/2(ERK1/2)、信号传导与转录激活子3(STAT3)蛋白活化的影响。方法:将体外培养A498细胞与不同浓度SAL(0、0.6、1.25、2.5、5、10、20、40、80、160、320μmol/L)孵育,计算半抑制浓度(IC),设置SAL 0、10、20和40μmol/L组。检测各组细胞侵袭细胞数,ELISA检测上清IL-6、IL-10、TNF-α水平,Western blot检测上皮细胞钙黏素(E-cad)、神经钙黏素(N-cad)、波形蛋白(Vimentin)、IL-10、TNF-α、ERK1/2、p-ERK1/2、STAT3和p-STAT3蛋白表达。结果:SAL对A498细胞增殖的抑制作用呈药物浓度依赖性,IC(23.41±2.45)μmol/L。SAL 20μmol/L组和SAL 40μmol/L组细胞存活率,侵袭细胞数明显低于SAL 0μmol/L组(P<0.05),细胞上清中IL-6和TNF-α水平低于SAL 0μmol/L组(P<0.05),IL-10水平高于SAL 0μmol/L组(P<0.05),细胞E-cad蛋白表达高于SAL0μmol/L组(P<0.05),N-cad、Vimentin、p-ERK1/2/ERK1/2和p-STAT3/STAT3蛋白表达低于SAL 0μmol/L组(P<0.05)。结论:SAL可以抑制A498细胞增殖和运动,其作用机制可能与抑制免疫炎症反应和ERK1/2和STAT3磷酸化有关。

Objective:To explore the effects of salidroside(SAL)on cell invasion,immune inflammatory cytokines,activation of extracellular signal-regulated kinase 1/2(ERK1/2)and signal transduction and transcriptional activators 3(STAT3)in renal carcinoma cells A498.Methods:A498 cells cultured in vitro were incubated with different concentrations of SAL(0,0.6,1.25,2.5,5,10,20,40,80,160,320 μmol/L). The 50% inhibiting concentration(IC)was calculated. SAL 0,10,20 and 40 μmol/L group were set up. The number of invaded cells in each group was detected. The levels of IL-6,IL-10 and TNF-α in supernatant were detected by ELISA. The expression of epithelial cadherin(E-cad),N-cadherin(N-cad),Vimentin(Vimentin),IL-10,TNF-α,ERK1/2,p-ERK1/2,STAT3 and p-STAT3 proteins were detected by by Western blot.Results:The inhibitory effects of SAL on the proliferation of A498 cells showed concentration-dependence of drug,with ICof(23.41±2.45)μmol/L. The cells survival rate and number of invaded cells in SAL 20 μmol/L group and SAL 40 μmol/L group were significantly lower than those in SAL 0 μmol/L group(P<0.05),levels of IL-6 and TNF-α in cell supernatant were lower than those in SAL 0 μmol/L group(P<0.05),IL-10 level was higher than that in SAL 0 μmol/L group(P<0.05),expression of E-cad protein was higher than that in SAL 0 μmol/L group(P<0.05),and expression of N-cad,Vimentin,p-ERK1/2/ERK1/2 and p-STAT3/STAT3 were lower than that in SAL 0 μmol/L group(P<0.05).Conclusion:SAL can inhibit the proliferation and movement of A498 cells,whose action mechanism may be related to inhibit immune inflammation and phosphorylations of ERK1/2 and STAT3.