Anoikis resistance is a hallmark of cancer,and relates to malignant phenotypes,including cell migration,epithelial-mesenchymal transformation (EMT),metastasis and cancer stem cell maintenance.Anoikis research has become a hot topic in cancer research,but little is known about the underlying mechanisms of anoikis and how it drives tumor invasion and metastasis.Here,we established an approach to screen anoikis-resistant cell by suspension-culturing MCF 10A human breast epithelial cells after transduction of pooled lentiviral shRNA library.The presence and quantity of shRNAs in anoikis-resistant ceils was confirmed by next generation sequencing and eventually,we obtained 7 anoikis resistance related candidate genes.Further PPI network analysis showed nuclear respiratory factor-1 (NRF1) was a hub protein for information transmission between candidate genes,which has the highest connectivity in the specific cell death and apoptosis related module.From another perspective,the results once again highlighted the importance of NRF1(P-value=1.8E-2) in transcriptional regulation.Overexpression of NRF1 in MCF 10A cells enhanced cell migration,invasion and endowed the cells anoikis-resistant capacity.Conversely,suppression of NRF1 decreased resistance to anoikis and reduced malignancy in MDA-MB-231 cells.Further experiments confirmed that NRF1 led to EMT in above biological processes.The results from our survival analysis indicated that,in clinical breast cancer specimens,overexpression ofNRF 1 was significantly associated with poor prognosis (P<0.05).