RNF8 promotes epithelial-mesenchymal transition of breast cancer cells.
Epithelial-mesenchymal transition (EMT) is a crucial step for solid tumor progression and plays an important role in cancer invasion and metastasis. RNF8 is an ubiquitin E3 ligase with RING domain, and plays essential roles in DNA damage response and cell cycle regulation. However the role of RNF8 in the pathogenesis of breast cancer is still unclear.



The expression of RNF8 was examined in different types of breast cell lines by Western Blotting. EMT associated markers were examined by Immunofluorescence and Western Blotting in MCF-7 when RNF8 was ectopically overexpressed, or in MDA-MB-231 when RNF8 was depleted. Transwell and wound healing assays were performed to assess the effect of RNF8 on cell mobility. The xenograft model was done with nude mice to investigate the role of RNF8 in tumor metastasis in vivo. Breast tissue arrays were used to examine the expression of RNF8 by immunohistochemistry. Kaplan-Meier survival analysis for the relationship between survival time and RNF8 signature in breast cancer was done with an online tool ( http://kmplot.com/analysis/ ).



RNF8 is overexpressed in highly metastatic breast cancer cell lines. Overexpression of RNF8 in MCF-7 significantly promoted EMT phenotypes and facilitated cell migration. On the contrary, silencing of RNF8 in MDA-MB-231 induced MET phenotypes and inhibited cell migration. Furthermore, we proved that these metastatic behavior promoting effects of RNF8 in breast cancer was associated with the inactivation of GSK-3β and activation of β-catenin signaling. With nude mice xenograft model, we found that shRNA mediated-downregulation of RNF8 reduced tumor metastasis in vivo. In addition, we found that RNF8 expression was higher in malignant breast cancer than that of the paired normal breast tissues, and was positively correlated with lymph node metastases and poor survival time.



RNF8 induces EMT in the breast cancer cells and promotes breast cancer metastasis, suggesting that RNF8 could be used as a potential therapeutic target for the prevention and treatment of breast cancer.
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China.
Jingyu
J
Kuang
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China. lily@bjmu.edu.cn.
Li
L
Li
Department of Pathology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China.
Limei
L
Guo
The Irma H. Russo, MD Breast Cancer Research Laboratory, Fox Chase Cancer Center-Temple University Health System, Philadelphia, PA, 19111, USA.
Yanrong
Y
Su
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China.
Yuxuan
Y
Wang
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China.
Yongjie
Y
Xu
Department of Breast Surgery, the First Hospital of Jilin University, Changchun, 130021, China.
Xiaozhen
X
Wang
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China.
Shucong
S
Meng
Lab of Molecular Imaging, Health Science Analysis Center, Peking University, Beijing, 100191, China.
Liandi
L
Lei
Lab of Molecular Imaging, Health Science Analysis Center, Peking University, Beijing, 100191, China.
Luzheng
L
Xu
Department of Cell Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, China. gzshao@bjmu.edu.cn.
Genze
G
Shao
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D015398:Signal Transduction; D016019:Survival Analysis; D044767:Ubiquitin-Protein Ligases; D015854:Up-Regulation; D051176:beta Catenin / Q000378:metabolism
C495270:CTNNB1 protein, human; D004268:DNA-Binding Proteins; C486521:RNF8 protein, human; D051176:beta Catenin; D044767:Ubiquitin-Protein Ligases; D000071679:Glycogen Synthase Kinase 3 beta
Breast cancer; EMT; Metastasis; RNF8