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目的观察柴胡皂甙d(saikosaponin-d,SSd)对人肝癌细胞STAT3、COX-2表达的影响,探讨其抗肿瘤作用可能的信号通路机制。方法体外培养人肝癌SMMC-7721细胞,分别经IL-6、AG490和SSd作用后,免疫细胞化学及免疫印迹方法检测STAT3、磷酸化STAT3(p-STAT3)以及COX-2蛋白表达的变化,并探讨其关系。结果免疫细胞化学结果显示,IL-6诱导组人肝癌细胞p-STAT3和COX-2蛋白的表达较空白对照组显著升高,而加入AG490或SSd后p-STAT3、COX-2蛋白表达较IL-6诱导组明显减弱,SSd对STAT3蛋白表达则无明显影响;免疫印迹结果亦显示出类似变化:经IL-6作用,人肝癌细胞p-STAT3和COX-2的表达明显增强,AG490或SSd作用的人肝癌细胞p-STAT3表达呈现剂量依赖性下降,COX-2蛋白表达亦出现相应下调。结论p-STAT3参与了人肝癌细胞COX-2表达调节,SSd可能通过抑制STAT3磷酸化下调COX-2的表达而发挥抗肿瘤作用。
Objective To investigate the effect of saikosaponin-d (SSd) on the expression of STAT3 and COX-2 in human hepatocellular carcinoma cells and to explore the possible signaling pathway of anti-tumor effect. Methods Human hepatocellular carcinoma SMMC-7721 cells were cultured in vitro and the changes of STAT3, p-STAT3 and COX-2 protein expression were detected by immunocytochemistry and Western blotting after IL-6, AG490 and SSd respectively Explore its relationship. Results The results of immunocytochemistry showed that the expressions of p-STAT3 and COX-2 in human hepatocarcinoma cells were significantly higher than those in blank control group, while the expressions of p-STAT3 and COX-2 protein were significantly higher than those in control group -6 induced group was significantly weakened, SSd had no significant effect on STAT3 protein expression; Western blot results also showed similar changes: the expression of p-STAT3 and COX-2 in human hepatocellular carcinoma cells was significantly increased by IL-6, AG490 or SSd The expression of p-STAT3 in human hepatocellular carcinoma cells decreased in a dose-dependent manner, and the expression of COX-2 protein also decreased accordingly. Conclusion p-STAT3 is involved in the regulation of COX-2 expression in human hepatocellular carcinoma cells. SSd may play an antitumor effect by inhibiting the phosphorylation of STAT3 and down-regulating the expression of COX-2.