FOXD1-dependent RalA-ANXA2-Src complex promotes CTC formation in breast cancer

Abstract

Background Early metastasis is a key factor contributing to poor breast cancer (BC) prognosis. Circulating tumor cells (CTCs) are regarded as the precursor cells of metastasis, which are ultimately responsible for the main cause of death in BC. However, to date molecular mechanisms underlying CTC formation in BC have been insufficiently defined. Methods RNA-seq was carried out in primary tissues from early-stage BC patients (with CTCs≥5 and CTCs = 0, respectively) and the validation study was conducted in untreated 80 BC patients. Multiple in vitro and in vivo models were used in functional studies. Luciferase reporter, ChIP-seq, CUT&Tag-seq, and GST-pulldown, etc. were utilized in mechanistic studies. CTCs were counted by the CanPatrol™ CTC classification system or LiquidBiospy™ microfluidic chips. ERK1/2 inhibitor SCH772984 was applied to in vivo treatment. Results Highly expressed FOXD1 of primary BC tissues was observed to be significantly associated with increased CTCs in BC patients, particularly in early BC patients. Overexpressing FOXD1 enhanced the migration capability of BC cells, CTC formation and BC metastasis, via facilitating epithelial-mesenchymal transition of tumor cells. Mechanistically, FOXD1 was discovered to induce RalA expression by directly bound to RalA promotor. Then, RalA formed a complex with ANXA2 and Src, promoting the interaction between ANXA2 and Src, thus increasing the phosphorylation (Tyr23) of ANXA2. Inhibiting RalA-GTP form attenuated the interaction between ANXA2 and Src. This cascade culminated in the activation of ERK1/2 signal that enhanced metastatic ability of BC cells. In addition, in vivo treatment with SCH772984, a specific inhibitor of ERK1/2, was used to dramatically inhibit the CTC formation and BC metastasis. Conclusion Here, we report a FOXD1-dependent RalA-ANXA2-Src complex that promotes CTC formation via activating ERK1/2 signal in BC. FOXD1 may serve as a prognostic factor in evaluation of BC metastasis risks. This signaling cascade is druggable and effective for overcoming CTC formation from the early stages of BC.

Annexin A2 combined with TTK accelerates esophageal cancer progression via the Akt/mTOR signaling pathway

“…In HCC and breast cancer, ANXA2 was found to interact with FBXW7 and MIEN2 respectively [ 40 , 41 ]. Besides, ANXA2 can participate in the formation of various compounds to promote drug resistance in breast cancer [ 42 – 44 ]. ANXA2 is also associated with several classical cancer-related pathways such as PI3K/Akt signaling, Akt/mTOR signaling, and NF-κB signaling pathways in lung cancer, gastric cancer, and GBM respectively [ 34 , 45 , 46 ].…”

Lysyl oxidase-like 4 promotes the invasiveness of triple-negative breast cancer cells by orchestrating the invasive machinery formed by annexin A2 and S100A11 on the cell surface

“…Thus, TNBC cells can accelerate their invasion with multiple mechanisms, even with the single molecule annexin A2 ( 29 – 31 ). It has been reported that the cell surface presentation of annexin A2 requires Src-mediated tyrosine phosphorylation, which is readily elevated in TNBC cells ( 32 – 34 ). The extracellularly secreted LOXL4 catalytically works to lock annexin A2 on the cell membrane surface by multimerization modification.…”

Lysine Acetylome of Breast Cancer-Derived Small Extracellular Vesicles Reveals Specific Acetylation Patterns for Metabolic Enzymes

“…The protein Annexin A2 has been explored as a prognostic marker because of the expression of this protein in various cancer cells [ 72 ] including oral squamous cell carcinoma [ 73 ] and prostate cancer [ 74 , 75 , 76 ]. It was found that Annexin A2 is highly expressed in breast tumor tissues [ 77 ] and that FOXD1-dependent RalA-ANXA2-Src complex promotes circulating tumor cell formation in BC [ 78 ]. Moreover, Annexin A5 (ANXA5) was found to promote prostate cancer stem cells, pancreatic adenocarcinoma, sarcoma and tumorigenesis and progression of BC [ 79 ].…”