The miR-141/neuropilin-1 axis is associated with the clinicopathology and contributes to the growth and metastasis of pancreatic cancer

Lixin Ma; Bo Zhai; Huaqiang Zhu; Weidong Li; Wenjing Jiang; Liwang Lei; Shujun Zhang; Haiquan Qiao; Xian Jiang; Xueying Sun

doi:10.1186/s12935-019-0963-2

The miR-141/neuropilin-1 axis is associated with the clinicopathology and contributes to the growth and metastasis of pancreatic cancer

Cancer Cell Int. 2019 Sep 27:19:248. doi: 10.1186/s12935-019-0963-2. eCollection 2019.

Authors

Lixin Ma¹, Bo Zhai^{2

3}, Huaqiang Zhu⁴, Weidong Li^{2

3}, Wenjing Jiang³, Liwang Lei¹, Shujun Zhang⁵, Haiquan Qiao¹, Xian Jiang^{1

3}, Xueying Sun³

Affiliations

¹ 1Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001 China.
² 2Department of General Surgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, 150001 China.
³ 3The Hepatosplenic Surgery Center, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001 China.
⁴ 4Department of Hepatobiliary Surgery, Shandong Provincial Hospital, Jinan, 250021 China.
⁵ 5Department of Pathology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, 150001 China.

Abstract

Background: Neuropilin-1 (NRP-1) is a non-tyrosine kinase receptor interacting with multiple signaling pathways that underpin the biological behavior and fate of cancer cells. However, in pancreatic cancer, the mechanisms underlying the function of NRP-1 in cell proliferation and metastasis and the involvement of regulatory upstream miRNAs remain unclear.

Methods: Potential miRNAs were mined by using multiple bioinformatics prediction tools and validated by luciferase assays. The expression of NRP-1 and miRNA-141 (miR-141) in pancreatic tissues and cells was examined by immunohistochemistry, immunoblotting and/or real-time RT-PCR. Stable transfected cells depleted of NRP-1 were generated, and regulatory effects of miR-141 were investigated by transfecting cells with miR-141 mimics and anti-miR-141. Assays of cell viability, proliferation, cell cycle distribution, transwell migration and cell scratch were employed. Xenograft tumor models were established to assess the effects of NRP-1 depletion on tumorigenesis and liver metastasis, and therapeutic effects of miR-141 on tumor growth. The role of miR-141/NRP-1 axis in regulating epithelial-mesenchymal transition (EMT) by co-interacting the TGF-β pathway was examined.

Results: In this study, of 12 candidate miRNAs identified, miR-141 showed the strongest ability to regulate NRP-1. In pancreatic cancer tissues and cells, the expression level of NRP-1 was negatively correlated with that of miR-141. NRP-1 was highly expressed in pancreatic cancer tissues compared with normal pancreatic tissues, and its expression levels were positively correlated with tumor grade, lymph metastasis and AJCC staging. NRP-1 depletion inhibited cell proliferation by inducing cell cycle arrest at the G0/G1 phase through upregulating p27 and downregulating cyclin E and cyclin-dependent kinase 2, and reduced cell migration by inhibiting EMT through upregulating E-cadherin and downregulating Snail and N-cadherin. Through downregulating NRP-1, miR-141 mimics showed a similar effect as NRP-1 depletion on cell proliferation and migration. NRP-1 depletion suppressed tumor growth and liver metastasis and miR-141 mimics inhibited the growth of established tumors in mice. NRP-1 depletion and/or miR-141 mimics inhibited the activation of the TGF-β pathway stimulated by TGF-β ligand.

Conclusions: The present results indicate that NRP-1 is negatively regulated by miR-141 and the miR-141/NRP-1 axis may serve as potentially valuable biomarkers and therapeutic targets for pancreatic cancer.

Keywords: Cellular signaling; Epithelial–mesenchymal transition; Metastasis; MicroRNA-141; Neuropilin-1; Pancreatic cancer; Proliferation; Transforming growth factor β.