Human rhomboid family-1 modulates clathrin coated vesicle-dependent pro-transforming growth factor α membrane trafficking to promote breast cancer progressionResearch in context
Jie Li,
Tai-Ran Bai,
Shan Gao,
Zhuan Zhou,
Xue-Mei Peng,
Li-Song Zhang,
Dao-Lei Dou,
Zhi-Song Zhang,
Lu-Yuan Li
Affiliations
Jie Li
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Tai-Ran Bai
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Shan Gao
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Zhuan Zhou
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.
Xue-Mei Peng
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Li-Song Zhang
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Dao-Lei Dou
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China
Zhi-Song Zhang
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China; Corresponding authors at: Nankai University, 94 Weijin Road, State Key Laboratory of Medicinal Chemical Biology 203, Tianjin 300071, China.
Lu-Yuan Li
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300071, China.; Collaborative Innovation Center for Biotherapy and School of Medicine, Nankai University, Tianjin, 300071, China; Corresponding authors at: Nankai University, 94 Weijin Road, State Key Laboratory of Medicinal Chemical Biology 203, Tianjin 300071, China.
Background: Epidermal growth factor receptor (EGFR) signalling is critical in epithelial cancer development. Human rhomboid family-1 (RHBDF1) facilitates the secretion of TGFα, an EGFR ligand, in breast cancer; however, the underlying mechanism remains unclear. We evaluated the role for RHBDF1 in clathrin-coated vesicle (CCV)-dependent pro-TGFα membrane trafficking in breast cancer cells upon stimulation by G-protein coupled receptor (GPCR) agonists. Methods: RHBDF1 was silenced in various breast cancer cells using shRNA. TGFα levels, subcellular localization, and secretion were evaluated using ELISA, immunofluorescent staining, and coimmunoprecipitation. Phosphorylation and expression of relevant proteins were measured by western blotting. RHBDF1-dependent cell viability and invasion were measured. Findings: RHBDF1 mediates GPCR agonist-induced EGFR phosphorylation by promoting TGFα secretion in various types of breast cancer cells. RHBDF1 not only mediates ADAM17-dependent shedding of TGFα, but is essential in membrane trafficking of pro-TGFα. RHBDF1 silencing results in blocking of clathrin uncoating from CCV, a crucial step for the plasma membrane release of pro-TGFα. Interaction of RHBDF1 with auxilin-2, a CCV protein, determines the recruitment of HSC70 to CCV to facilitate clathrin uncoating. RHBDF1 function is required for the proliferation and mobility of breast cancer cells upon stimulation by Sphingosine 1 Phosphate (S1P), a GPCR agonist. We demonstrate a significant correlation between RHBDF1 overexpression and EGFR activation in breast cancer tissues. Interpretation: RHBDF1 is an indispensable component of the protein trafficking machinery involved in GPCR-mediated EGFR transactivation, and is an attractive therapeutic target for cancer. Fund: National Natural Science Foundation of China (81,672,740 to ZSZ, 81,272,356 and 81,330,029 to LYL). Keywords: RHBDF1, EGFR, TGFα, Membrane trafficking, Clathrin-coated vesicle, Breast cancer, Progression
