黄芪注射液对人子宫内膜癌细胞β-catenin和E-cadherin表达的影响

Effects of astragalus injection on the expression of β-Catenin and E-cadherin in human endometrial carcinoma cells

目的研究黄芪注射液对子人宫内膜癌HEC-1-B细胞β-catenin和E-cadherin基因和蛋白表达的影响。方法分别采用RT-PCR和免疫组化的方法检测β-catenin和E-cadherin基因和蛋白的表达。用含不同浓度的黄芪注射液的条件培养基分别培养人子宫内膜癌细胞株HEC-1-B 24、48、72 h后,应用RT-PCR检测各组细胞中β-catenin、E-cadherin基因表达,应用免疫细胞化学法检测HEC-1-B中β-catenin、E-cadherin蛋白表达。结果 RT-PCR结果显示,随着子宫内膜癌细胞在含有黄芪注射液的条件培养基中培养时间的延长,细胞中E-cadherin基因表达逐步增强(P<0.05),而β-catenin基因表达逐步减弱(P<0.05)。免疫组化染色结果显示,与对照组比较,黄芪注射液可明显降低子宫内膜癌HEC-1-B细胞中β-catenin蛋白的表达、增加E-cadherin蛋白的表达(P<0.05),而且黄芪注射液浓度越高处理时间越长β-catenin表达更低(P<0.05),黄芪注射液浓度越高E-cadherin表达更高(P<0.05)。结论黄芪注射液可呈剂量和时间依赖性促进E-cadherin基因和蛋白的表达、抑制β-catenin基因和蛋白的表达,从而使WNT信号通路受到抑制。这可能是其发挥抗肿瘤作用的机制之一。

Objective To investigate the effects of astragalus injection on the m RNA and protein expression of β-catenin and E-cadherin in HEC-1-Bendometrial carcinoma cells.Methods Cell climbing slices of HEC-1-B cells were prepared for the intervening experiment of astragalus injection on endometrial carcinoma cells,then their m RNA and protein expression ofβ-catenin and E-cadherin detected by RT-PCR and immunohistochemistry.Results RT-PCR results showed that with the incubation time of endometrial cancer cells in conditioned medium containing astragalus injection increased,E-cadherin gene expression of the cells gradually increased（P0.05）,while the β-catenin gene expression gradually decreased（P0.05）.The immunocytochemistry result indicated thatastragalus could significantly decrease β-catenin expression and increase E-cadherin expression in HEC-1-B cells（P0.05）.Moreover,accompanied by elevated concentration and treatment time of astragalus injection,β-catenin expression dropped progressively（P0.05）,meanwhile with the concentration of astragalus injection raised,E-cadherin expression gradually increased（P0.05）.Conclusion Astragalus injection could significantly inhibit WNT signaling pathway by promoting m RNA andprotein expression of E-cadherin,decreasing m RNA and protein expression ofβ-cateninin a dose- and time-dependent manner,which might be one of the mechanisms for its anti-tumor effect.