SOCS1调控JAK/STAT通路传导抑制肝癌细胞增殖、侵袭、迁移的机制研究

SOCS1 Inhibits the Proliferation,Invasion and Migration of Hepatoma Cell by Regulating the JAK/STAT Signaling Pathway

目的探讨SOCS1在肝癌进展过程中的功能及其分子机制。方法采用qRT-PCR和Western bolt检测人正常肝细胞和肝癌细胞系中SOCS1的表达。构建SOCS1过表达及干扰表达载体,转染肝癌细胞,MTT检测肝癌细胞增殖,细胞划痕实验检测细胞迁移情况,transwell实验检测细胞侵袭性变化。qRT-PCR检测JAK2、STAT3 mRNA水平;Western bolt检测JAK2、STAT3蛋白和磷酸化水平。通过SOCS1 siRNA和JAK/STAT通路抑制剂氯硝柳胺共处理肝癌细胞,观察肝癌细胞增殖、迁移及侵袭的变化。结果SOCS1在肝癌细胞系中显著低表达,SOCS1过表达后肝癌细胞增殖活性显著下调,细胞迁移能力显著下降,1/2划痕愈合时间增长,细胞侵袭能力显著下降,而SOCS1干扰表达则促进细胞增殖、迁移和侵袭。SOCS1表达变化不影响JAK2、STAT3 mRNA和蛋白表达水平,但SOCS1过表达抑制JAK2、STAT3磷酸化,SOCS1干扰表达则增强JAK2、STAT3磷酸化水平。JAK/STAT通路抑制剂氯硝柳胺能够明显恢复SOCS1干扰表达对肝癌细胞增殖、迁移和侵袭性的影响。结论SOCS1通过调控JAK/STAT信号通路活性,进而抑制肝癌细胞增殖、迁移和侵袭。

Objective To explore the functional role and specific molecular mechanism of SOCS1 in the progression of liver cancer.Methods The expression of SOCS1 in normal hepatocytes and hepatoma cells was detected by qRT-PCR and Western blot.The overexpression and interference expression vectors of SOCS1 were constructed and transfected into hepatoma cells.Cell proliferation was detected by MTT,cell migration was detected by cell scratch assay,and the changes of cell invasion were detected by transwell assay.qRT-PCR was used to detect the expression of JAK2 and STAT3 mRNA,and Western bolt was used to detect protein expression levels and phosphorylation levels of JAK2 and STAT3.Hepatoma cells were co-treated with SOCS1 siRNA and JAK/STAT pathway inhibitor niclosamide to verify that SOCS1 regulated hepatoma cell proliferation,migration and invasion through JAK/STAT signaling pathway.Results Compared with normal hepatocytes,SOCS1 was significantly lower expressed in hepatoma cell lines.The proliferation activity of hepatoma cells was significantly down-regulated,the cell migration ability was significantly decreased,healing time of 1/2 scratches was increased,and the cell invasion ability was significantly decreased after overexpression of SOCS1,while the interference expression of SOCS1 promoted cell proliferation,migration and invasion.SOCS1 expression changes did not affect JAK2,STAT3 mRNA and protein expression levels,but SOCS1 overexpression inhibited JAK2 and STAT3 phosphorylation,and SOCS1 interference expression promoted JAK2,STAT3 phosphorylation.Compared with si-SOCS1 treating hepatoma cells alone,the JAK/STAT pathway inhibitor niclosamide restored the proliferation,migration and invasiveness of hepatoma cells caused by the interference expression of SOCS1.Conclusion SOCS1 inhibits the activity of JAK/STAT signaling pathway,thereby inhibiting the proliferation,migration and invasion of hepatoma cells.