C-Myc promotes lymphatic metastasis of pancreatic neuroendocrine tumor through VEGFC upregulation.

Tsung‐Ming Chang,Li‐Tzong Chen,Yan‐Shen Shan,Hui‐You Lin,Wen‐Chun Hung,Kuo‐Wei Huang,Pei‐Yi Chu,Jeffrey S Chang,Hui‐Jen Tsai

doi:10.1111/cas.14717

Abstract

Pancreatic neuroendocrine tumor (pNET) is a pancreatic neoplasm with neuroendocrine differentiation. pNET in early stage can be treated with surgical resection with long‐term survival, whereas the prognosis of pNET with locoregional or distant metastasis is relatively poor. Lymphangiogenesis is essential for tumor metastasis via the lymphatic system and may overhead distant metastasis. c‐Myc overexpression is involved in tumorigenesis. The role of c‐Myc in lymphangiogenesis is unclear. In this study, we evaluated the mechanism and effect of c‐Myc on lymphangiogenesis of pNET via interaction of lymphatic endothelial cells (LECs) and pNET cells. Lymph node metastasis was evaluated in pNET xenograft mice. Potential target agents to inhibit lymph node metastasis were evaluated in an animal model. We found that vascular endothelial growth factor C (VEGFC) expression and secretion was increased in pNET cell lines with c‐Myc overexpression. c‐Myc transcriptionally upregulates VEGFC expression and the secretion of pNET cells by directly binding to the E‐box of the VEGFC promoter and enhances VEGF receptor 3 phosphorylation and the tube formation of LECs. c‐Myc overexpression is associated with lymph node metastasis in pNET xenograft mice. Combinational treatment with an mTOR inhibitor and c‐Myc inhibitor or VEGFC‐neutralizing chimera protein reduced lymph node metastasis in the mice with c‐Myc overexpression. The mTOR inhibitor acts on lymphangiogenesis by reducing VEGFC expression in pNET cells and inhibiting the tube formation of LECs. In conclusion, mTOR and c‐Myc are important for lymphangiogenesis of pNET and are potential therapeutic targets for prevention and treatment of lymph node metastasis in pNET.

Highlights

We demonstrate that c-Myc overexpression transcriptionally upregulates vascular endothelial growth factor C (VEGFC) expression in Pancreatic neuroendocrine tumor (pNET) cells, which promotes lymphangiogenesis of pNET cells in vitro and in vivo
We have shown that c-Myc promoted tumor growth and lymph node metastasis in QGP-1 xenograft mice
Activation of mTOR pathway was shown to be associated with increased lymphatic vessel density (LVD) and lymph node metastasis via upregulation of VEGFC in various cancers in in vitro and in vivo models and in human samples.[45,46,47] mTOR inhibitors decreased VEGFC/D expression and diminished lymphangiogenesis and lymph node metastasis in various cancer models.[45,46,47,48]

Summary

| INTRODUCTION

Pancreatic neuroendocrine tumor (pNET) is a rare histologic subtype of pancreatic cancer with the expression of neuroendocrine markers. QGP-1, a human pNET cell line, was purchased from the Japanese Collection of Research Bioresources (JCRB, Tokyo, Japan). NIT-1, a mouse pNET cell line, was purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). Human dermal lymphoepithelial cells (HDLECs), which was kindly provided by Dr Wen-Chun Hung (National Health Research Institutes, Tainan, Taiwan), were obtained from Promo Cell and cultured in endothelial cell growth medium MV2 (EGM-MV2) according to manufacturer's instruction. The passage of these cell lines used in this study was

| MATERIALS AND METHODS

Findings

| DISCUSSION