| Objective: To explore the mechanism of oxidized ATM on invasion and metastasis of hypoxic breast cancer cell via glucose metabolism.Methods: Oxidized ATM was assessed by western blotting and immunofluorescence;PFKP and CS expression by quantitative real-time polymerase chain reaction(qRT-PCR),western blotting and immunohistochemistry;migration and invasion analysis by transwell assays in vitro,and orthotopic Xenografts in vivo.The mechanism of oxidized ATM-mediated tumor metastasis was analyzed via shRNA,siRNA and overexpression.The interaction between UBR5 and CS was evaluated by IP.The regulation of HIF1 A to PFKP transcriptionally was tested by ChIP and luciferase assay.Results: oxidized ATM enhanced intracellular citrate via promoting PFKP and CS.PFKP is transcriptionally regulated by HIF1 A and CS is post-transcriptional regulation via UBR5-mediated ubiquitination,respectively.Accumulation of citrate in cytoplasm or exogenous citrate significantly enhances cell migration,invasion and metastasis of hypoxic TNBC cells in vitro and in mice xenografts.Conclusions: These results unravel a novel function of oxidized ATM in promoting migration,invasion and metastasis of breast cancer. |
