白杨素对肝癌干样细胞球形成及CK2α和Gli1表达的影响

Effects of Chrysin on Expression of CK2α and Gli1 and Spheroid Formation in Liver Cancer Stem-like Cells from SMMC-7721 Cell Line

本文研究了中药蜂胶胶囊主要活性成分白杨素对人肝癌细胞系SMMC-7721肝癌干样细胞(liver cancer stem-like cells,LCSLCs)球形成抑制作用及其机制。采用干细胞条件培养基悬浮培养SMMC-7721细胞得到第3代球细胞定义为(SFCs)。通过球形成试验评价白杨素对SFCs球形成率的影响,Western blot分析白杨素处理SFCs的CD133、CD44、CK2α和Gli1蛋白表达水平,CK2α siRNA转染SFCs探讨白杨素的作用机制。结果表明不同浓度白杨素降低SFCs球形成率,下调CD133、CD44、CK2α和Gli1蛋白表达。与乱序对照siRNA转染细胞相比,CK2α siRNA转染下调CK2α和Gli1蛋白表达,同时,协作增强白杨素抑制CK2α和Gli1蛋白表达作用。说明白杨素可能通过抑制CK2α表达阻断Hedgehog信号转导从而抑制SFCs球形成。

To investigate the inhibitory effect of chrysin from traditional Chinese medicine propolis capsule on sphere forming capability of liver cancer stem-like cells derived from human hepatocelluiar carcinoma SMMC-7721 ccll line and its mechanism. Suspension culture with the stem ceil-condition medium was used to obtain the third generation spheroids derived from SMMC-7721 cell line defined as SFCs. Sphere formation assay was used to evaluate the effects of chrysin on sphere forming rate of SFCs. Western blot was used to analyze the expression levels of CD133, CD44, CK2α and Glil proteins in SFCs treated with chrysin. CK2αsiRNA transfection was employed to approach the mechanism of chrysin action. The resuh showed that chrysin reduced the sphere forming rate and down-regulated expressions of CD133, CD44, CK2α and Glil proteins of SFCs. Compared with SFCs transfected with scrambled control siRNA, a decrease of CK2α and Glil protein expression level in SFCs transfeeted with CK2αsiRNA was observed. In addition, CK2αsiRNA transfec- tion cooperatively enhanced chrysin down-regulated the expression of CK2α and Glil proteins in SFCs. It was showed that chrysin inhibited the sphere forming capability of SFCs may down-regulate CK2α expression through blocking Hedgehog signaling.