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目的研究热休克同源蛋白73(heat shock cognate protein73000,Hsc73)在基质细胞衍生因子1(SDF-1)/趋化因子CXC受体4(CXCR4)轴促进肾癌转移中的作用,确定Hsc73是否参与了CXCR4的核定位。方法通过蛋白电泳观察CXCR4高表达后Hsc73在A498细胞中的表达情况。通过免疫染色、核蛋白免疫共沉淀等方法观察A498细胞在经过SDF-1刺激后Hsc73在细胞内表达情况,以及Hsc73与CXCR4结合情况。结果 CXCR4高表达后,A498细胞中Hsc73表达明显增高。Hsc73主要表达在A498细胞胞质中,在SDF-1刺激后出现部分转移至细胞核。提取SDF-1刺激后的A498细胞核蛋白进行CXCR4免疫共沉淀,其中发现了Hsc73。结论 Hsc73作为细胞内常见的分子伴侣蛋白,参与了CXCR4在细胞中的转运。对于SDF-1/CXCR4信号通路激活的部分环节起到了关键作用,并可能参与了CXCR4核转位的过程。
Objective To investigate the role of heat shock cognate protein 73000 (Hsc73) in the promotion of renal cell carcinoma metastasis by stromal cell-derived factor 1 (SDF-1) / chemokine CXC receptor 4 (CXCR4) axis and to determine whether Hsc73 Participated in the nuclear localization of CXCR4. Methods The expression of Hsc73 in A498 cells was observed by protein electrophoresis after high expression of CXCR4. The expression of Hsc73 in A498 cells after SDF-1 stimulation and the binding of Hsc73 to CXCR4 were observed by immunostaining and nuclear co-immunoprecipitation. Results After CXCR4 was overexpressed, the expression of Hsc73 in A498 cells was significantly increased. Hsc73 is mainly expressed in the cytoplasm of A498 cells, and partially translocates to the nucleus after SDF-1 stimulation. CXCR4 co-immunoprecipitation was performed on the A498 cell nucleoprotein extracted from SDF-1 cells, in which Hsc73 was found. Conclusion Hsc73, as a common molecular chaperone in cells, is involved in the transport of CXCR4 in cells. It plays a key role in the part of the activation of SDF-1 / CXCR4 signaling pathway and may be involved in the process of CXCR4 nuclear translocation.