Key Laboratory of Tumor Metastasis of Liaoning Province, Department of Pathology, Dalian Medical University, Dalian 116044, China.
Key Laboratory of Tumor Metastasis of Liaoning Province, Department of Pathology, Dalian Medical University, Dalian 116044, China.
Biochem Biophys Res Commun. 2014 Feb 28;445(1):58-63. doi: 10.1016/j.bbrc.2014.01.119. Epub 2014 Feb 1.
We report for the first time that Guanine nucleotide-binding protein subunit beta-2-like 1 (RACK1) formed a complex with Annexin A7. Hca-F and Hca-P are a pair of syngeneic mouse hepatocarcinoma cell lines established and maintained in our laboratory. Our previous study showed that both Annexin A7 and RACK1 were expressed higher in Hca-F (lymph node metastasis >70%) than Hca-P (lymph node metastasis <30%). Suppression of Annexin A7 expression in Hca-F cells induced decreased migration and invasion ability. In this study, knockdown of RACK1 by RNA interference (RNAi) had the same impact on metastasis potential of Hca-F cells as Annexin A7 down-regulation. Furthermore, by co-immunoprecipitation and double immunofluorescence confocal imaging, we found that RACK1 was in complex with Annexin A7 in control cells, but not in the RACK1-down-regulated cells, indicating the abolishment of RACK1-Annexin A7 interaction in Hca-F cells by RACK1 RNAi. Taken together, these results suggest that RACK1-Annexin A7 interaction may be one of the means by which RACK1 and Annexin A7 influence the metastasis potential of mouse hepatocarcinoma cells in vitro.
我们首次报道鸟嘌呤核苷酸结合蛋白亚基β-2 样 1(RACK1)与膜联蛋白 A7 形成复合物。Hca-F 和 Hca-P 是我们实验室建立和维持的一对同源鼠肝癌细胞系。我们之前的研究表明,膜联蛋白 A7 和 RACK1 在 Hca-F(淋巴结转移>70%)中的表达均高于 Hca-P(淋巴结转移<30%)。在 Hca-F 细胞中抑制膜联蛋白 A7 的表达会导致迁移和侵袭能力下降。在这项研究中,通过 RNA 干扰(RNAi)敲低 RACK1 对 Hca-F 细胞转移潜能的影响与下调膜联蛋白 A7 的影响相同。此外,通过共免疫沉淀和双免疫荧光共聚焦成像,我们发现 RACK1 在对照细胞中与膜联蛋白 A7 形成复合物,但在 RACK1 下调的细胞中没有,这表明 RACK1 RNAi 使 Hca-F 细胞中 RACK1-膜联蛋白 A7 相互作用被消除。综上所述,这些结果表明,RACK1-膜联蛋白 A7 相互作用可能是 RACK1 和膜联蛋白 A7 影响体外鼠肝癌细胞转移潜能的一种方式。