TGF-β1 promotes human breast cancer angiogenesis and malignant behavior by regulating endothelial-mesenchymal transition

Zi-Xiong Li; Jie-Xin Chen; Ze-Jun Zheng; Wang-Jing Cai; Xiong-Bin Yang; Yuan-Yuan Huang; Yao Gong; Feng Xu; Yong-Song Chen; Ling Lin

doi:10.3389/fonc.2022.1051148

TGF-β1 promotes human breast cancer angiogenesis and malignant behavior by regulating endothelial-mesenchymal transition

Front Oncol. 2022 Nov 16:12:1051148. doi: 10.3389/fonc.2022.1051148. eCollection 2022.

Authors

Zi-Xiong Li^{1

2}, Jie-Xin Chen¹, Ze-Jun Zheng¹, Wang-Jing Cai¹, Xiong-Bin Yang¹, Yuan-Yuan Huang¹, Yao Gong³, Feng Xu⁴, Yong-Song Chen⁵, Ling Lin^{1

3}

Affiliations

¹ Department of Rheumatology and Immunology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
² Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
³ Department of Rheumatology, Shantou University Medical College, Shantou, China.
⁴ Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
⁵ Department of Endocrinology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.

Abstract

Background: Endothelial-mesenchymal transition (EndMT) is an important process of angiogenesis, which plays a significant role in in tumor invasion and metastasis, while its regulatory mechanisms in breast cancer remain to be fully elucidated. We previously demonstrated that tumor-associated macrophages (TAMs) can induce EndMT in endothelial cells by secreting CCL18 through the activation of the TGF-β and Notch signaling pathways in breast cancer. This study was designed to study the role of EndMT in breast cancer angiogenesis and progression in order to explore the underlying mechanism.

Methods: Immunohistochemistry (IHC) was used to evaluate the expression of microvascular density (MVD) and EndMT markers in breast cancer. TGF-β1 was used to induce EndMT models of differentiated-endothelial breast cancer stem-like cells (BCSLCs). In vitro cell migration, proliferation and matrigel tube-formation assays, as well as in vivo nude mouse tumor-bearing model and nude mouse dorsal skinfold window chamber (DSWC) model, were utilized to investigate the effects in order to explore the mechanism of EndMT induced by TGF-β1 on breast cancer progression.

Results: In this study, we demonstrated that the EndMT markers were positively associated with MVD indicating unfavorable prognosis of invasive ductal carcinoma (IDC) patients. Functionally, TGF-β1 promoted migration, proliferation and angiogenesis of differentiated-endothelial BCSLCs by inducing EndMT in vitro and promoted tumor growth and angiogenesis in vivo. Mechanically, we revealed TGF-β1 induced EndMT by activation of TGF-β and Notch signaling pathways with increase of p-Smad2/3 and Notch1 expression. Moreover, we found Snail and Slug were key factors of TGF-β and Notch signaling pathways.

Conclusion: Our findings elucidated the mechanism of TGF-β1 in the promotion of angiogenesis and progression by EndMT in breast cancer.

Keywords: BCSLC; EndMT; TGF-β1; angiogenesis; breast cancer; dorsal skinfold window chamber.