目的本研究旨在探讨Wnt/β-catenin信号通路和葡萄糖调节蛋白78(glucose_regulated protein 78,GRP78)两者在肠癌细胞中的调控关系。方法本研究采用氯化锂(LiC1)处理肠癌HT~29和DLD1细胞,激活其Wnt/β-catenin信号通路。通过Western blot和荧光定最PCR,检测Wnt信号活化对GRP78表达的影响;通过对细胞组分的分离和免疫荧光染色技术分析wnt信号活化对GRP78细胞膜转运的影响。结果LiCl处理能够显著激活HT-29和DLD1细胞中的Wnt/β-catenin信号通路。GRP78蛋白在HT-29细胞膜表面呈独特的“簇状”分布,LiCI处理明显提高了细胞中GRP78的转录和翻译水平,且GRP78在细胞表面的分布水平也有明显增加。结论肠癌细胞中GRP78的表达和细胞膜转位受Wnt/β-catenin信号通路活性的调控。
Objective To investigate the regulation of Wnt/β-catenin signaling pathway and glucose-regulated protein 78 (GRF78) in colon cancer. Methods To activate the Wnt/β-catenin signaling pathway, HT-29, DLD1 were treated with different concentrations of LiC1. The expression of GRP78 mRNA and protein were analyzed by QT-PCR and Western blot assay respectively. Separations sub-cellular components and immunofluorescence were performed to explore the membrane translocation of the GRP78. Results The Wnt signaling pathway was activated by LiC1 treatment which raised the levels of GRP78 transcription and translation in eolorectal cancer. In addition, the distribution of cell surface GRF78 presented in unique "clusters" forms. The distribution of cell surface GRP78 levels was also significantly increased by LiC1 treatment. Conclusions Expression and membrane translocation of the GRP78 can be regulated by Wnt/β-catenin signaling activity.