Cxcl-1 Derived From Tumor-associated Macrophages Promotes Breast Cancer Metastasis Via NF/?B/SOX4 Signaling

ObjectiveInvestigate the effects of TAMs-derived CXCL1 in breast cancer metastasis and the role of XIAOPI fomula in breast cancer metastasis,which might become a novel strategy for breast cancer metastasis prevention and treatment.Methods1.The cytokines of high expression of TAMs cells were screened by Cytokine array.And we applied the immunohistochemical technique to analyze the distribution of cxcl-1 and EMT in MMTV-Pyvt transgenic tumor mice.2.To explore the impact of cxcl-1 on the breast cancer cells proliferation,migration and invasion through MMT,scratch assay,transwell,gelatinase spectrum experiment and Western blot.3.Using QPCR to screen the highest responsive gene following CXCL1 administration.Then we tranfected the relevant gene cDNA into MCF-7,MDA-MB-231 and inhibited the relevant gene by delivering siRNA or sh RNA.Simultaneously,we uesed Western blot to detect the expression of signaling pathways related proteins.4.In vivo,we observed the effects of TAMs-cxcl-1 in the lung metastasis and tumor growth of Nod-scid mice by vivo imaging instrument and the expression of the related proteins by immunohistochemical and immunofluorescence technique.5.The relationship between cxcl-1 and survival of breast cancer patients was verified by human breast cancer tissue microarray.6.The vital target of xiaoyi particle was verified by the Cytokine array experiment.7.The scratch test and transwcll test proved that XIAOPI fomula could inhibit the ability of breast cancer metastatic invasion.Resultscytokine array validated that CXCL1 was the most-abundant chemokine secreted by TAMs,and TAMs-derived CXCL1 could promote breast cancer migration and invasion ability,as well as epithelial-mesenchymal transition(EMT)process in both mouse and human breast cancer cells.QPCR screening further validated SOX4 as the highest responsive gene following CXCL1 administration.Mechanistic study revealed that CXCL1 activates SOX4 transcription via NF-?B pathway.In vivo breast cancer xenografts demonstrated that CXCL1 silencing in TAMs resulted in a significant reduction in breast cancer growth and metastatic burden.Bioinformatic analysis and clinical investigation finally suggested that high CXCL1 expression was significantly correlated with breast cancer lymph node metastasis,poor overall survival and basal-like.TAMs/CXCL1 is the most important target of the XIAOPI fomula in breast metastasis.ConclusionTAMs/CXCL1 promotes breast cancer metastasis via NF-?B/sox4 activation.The XIAOPI fomula can inhibit the ability of breast cancer metastatic invasion through TAMs/CXCL1 target.