Metastasis-associated in colon cancer-1 upregulates vascular endothelial growth factor-C/D to promote lymphangiogenesis in human gastric cancer

doi:10.1016/j.canlet.2014.11.035

Cancer Letters

Volume 357, Issue 1, 1 February 2015, Pages 242-253

https://doi.org/10.1016/j.canlet.2014.11.035 Get rights and content

Highlights

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Lymphangiogenesis is actively contributed to tumor lymphatic metastasis.
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Metastasis-associated in colon cancer-1 (MACC1) induces gastric cancer lymphangiogenesis.
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MACC1 upregulates VEGF-C/VEGF-D expression to promote lymphangiogenesis.
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MACC1 induced VEGF-C/VEGF-D upregulation is regulated by c-Met signaling.
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MACC1 may act as a new target for lymphangiogenesis in gastric cancer.

Abstract

Lymphangiogenesis is actively contributed to lymphatic metastasis in gastric cancer (GC), and vascular endothelial growth factor (VEGF)-C and VEGF-D are key regulators for lymphangiogenesis. Metastasis-associated in colon cancer-1 (MACC1) was reported to be associated with lymph node metastasis in a few clinical studies, while little is known about the role of MACC1 in lymphangiogenesis. Hence, in the present study, we explored the potential role of MACC1 in lymphangiogenesis as well as the underlying mechanisms. By clinical observation, we found a positive relationship between MACC1 and lymphangiogenesis. Besides, similar results were also obtained from in vivo and in vitro studies. With an indirect co-culture system, we got that supernatant from MACC1 overexpressed GC cells accelerated human lymphatic endothelial cells' (HLECs') capacity of tube-like formation through enhancing cell proliferation and migration. Moreover, MACC1 overexpressed xenografts also presented more lymphatic vessels. Furthermore, MACC1 significantly increased the expression of VEGF-C/VEGF-D in GC cells and transplanted tumors, which was subsequently suppressed by c-Met inhibitor. All these data suggested a critical role for MACC1 in lymphatic dissemination of GC, providing evidence that MACC1 upregulated VEGF-C/VEGF-D secretion to promote lymphangiogenesis via c-Met signaling.

Introduction

Lymph nodes are the first metastasis destination for gastric cancer (GC) cells, and lymphatic vessels serve as an important route for the spread of cancer cells [1]. Recent experimental and clinical data have suggested that lymphangiogenesis actively contributes to GC metastasis based on the observations that lymphatic vessel density is correlated with the extent of lymph node metastasis and/or poor prognosis [2], [3]. Moreover, some animal tumor models have revealed that expression of lymphangiogenic growth factors leads to formation of lymphatic vessels and that lymphangiogenesis is accompanied by enhanced lymphatic metastasis [2]. These findings strongly suggested a crucial role of lymphangiogenesis in lymph node metastasis, and inhibition of lymphangiogenesis may largely contribute to blocking tumor metastasis. Therefore, more and more studies have focused on lymphangiogenesis and the correlated molecular mechanisms [4].

Metastasis-associated in colon cancer-1 (MACC1) was reported to be overexpressed in several tumors and mainly contributed to tumor metastasis [5], [6], [7], [8]. Several studies supported MACC1 as a new remarkable biomarker for disease prognosis and a promising therapeutic target for anti-tumor and anti-metastatic in a variety of solid cancers based on the facts that MACC1 promotes tumor cell proliferation and migration [5], [6], [7], [9]. Our previous studies also confirmed that MACC1 enhances GC cell migration and invasiveness through facilitating pseudopodia formation and promoting epithelial mesenchymal transition [6], which was considered to be the first step of tumor metastasis. Furthermore, clinical studies showed that MACC1 is associated with lymph node metastasis and vascular invasion in several solid tumors [10], [11]. These studies strongly suggested that MACC1 mainly contributes to tumor metastasis by accelerating cell motility and promoting spread of cancer cells. While, previous studies only demonstrate the mechanism of MACC1-induced cell motility, whether MACC1 were contributed to the spread of cancer cells and especially which routes were affected by MACC1 remains unclear. Therefore, combined with the correlation of MACC1 and lymph node metastasis, we focused on the role of MACC1 in lymphatic metastasis and tried to find out whether MACC1 upregulates tumor-induced lymphangiogenesis to further promote tumor metastasis.

As the best known lymphangiogenic signaling system, vascular endothelial growth factor-C (VEGF-C)/VEGF-D and VEGF receptor 3 (VEGFR-3) signaling axis, has been proven to play an important role in lymphangiogenesis in cellular and animal models [12], [13]. As members of vascular endothelial growth factor family, VEGF-C/VEGF-D has been reported to be secreted or facilitated by tumor cells and peritumoral stroma associated proteins [like matrix metalloproteinase (MMPs) and fibronectin] [14], [15]. Interestingly, our previous study has proven that MACC1 upregulates the expression of extracellular matrix related components including fibronectin, MMP2 and MMP9 in GC cells [6], which were also important for VEGF-C/VEGF-D induced lymphangiogenesis. Besides, binding sites for SP1 are considered to be the putative promoter of VEGF-C [16]. Studies also found that MACC1 could bind with SP1 sites directly or indirectly [17], which strongly suggested a promoting role of MACC1 in regulating VEGF-C. However, the actual effects and mechanisms of MACC1 on VEGF-C and VEGF-D in GC still remain inapprehensive. Additionally, as a key downstream signaling of MACC1, hepatocyte growth factor (HGF)/c-Met signaling axis was reported to be associated with lymphangiogenesis in tumor tissues by clinical and experimental studies [18], [19], [20], [21], while little is known about the relationship between HGF/c-Met and VEGF-C/VEGF-D.

Therefore, to examine whether MACC1 is involved in lymphatic metastasis, we evaluated the expression of MACC1 and lymph node metastasis as well as lymphatic vessel density in human gastric tumor tissues. Besides, in vivo and in vitro experiments were also performed to identify the roles of MACC1 in lymphangiogenesis. Our hypothesis was that MACC1 upregulated the expression of VEGF-C and VEGF-D to promote lymphangiogenesis in GC, which was affected by HGF/c-Met signaling pathway.

Section snippets

Study population and tissue samples

All tissue samples investigated in the study were obtained from patients who underwent curative gastrectomy (I–III stage, n = 151) or palliative surgery (IV stage, n = 39) from 2005 to 2012 at Division of Pathology, Nanfang Hospital, Guangzhou, China. All these cases were historically diagnosed at Nanfang Hospital. Besides, the corresponding clinicopathologic classification and staging of these cases were determined according to the 7th edition proposed by the American Joint Committee on Cancer

Expression of MACC1 was correlated with LVD in human gastric cancer tissues

In order to clarify the potential relationship between MACC1 and lymphangiogenesis, 190 gastric cancer tissues were obtained and immunostained with MACC1 and LYVE-1 (lymphatic vessel marker). Accordingly, MACC1 were predominantly found in cytoplasm of cancer cells (Fig. 1A, first line) and occasionally expressed in nucleus of cells (data not shown). Among them, 78.9% of the patients presented with an overexpression of MACC1, while 21.1% patients with a less expression. LYVE-1-positive vessels

Discussion

The present study mainly talked about the following issues: 1) we first gave the evidence that MACC1 was associated with lymphatic vessel metastasis as well as poor prognosis of GC; 2) MACC1 upregulated VEGF-C/VEGF-D to promote lymphangiogenesis in vivo and in vitro; 3) HGF/c-MET signaling pathway was involved in the MACC1-mediated lymphangiogenesis.

Extent of lymph node metastasis critically determines the clinical outcome of GC [31]. Lymphangiogenesis is a primary cause for lymph node

Conclusions

In summary, the present study showed a new potential molecular mechanism on lymphangiogenesis that MACC1 regulates expression of VEGF-C/VEGF-D to further affect lymphangiogenesis, which is influenced by HGF/c-Met signaling pathway. The findings also suggested a new role of MACC1 in GC metastasis and further proved the potential application of MACC1 in GC therapy.

Financial support

This work was supported by grants from the National Natural Science Foundation of China (31271564 to W.L., 81472314 to W.L. and 81302155 to N.H.); and the Special Foundation for National Clinical Specialties of China (to Department of Oncology Nanfang Hospital); and Guangzhou Science and Technology Plan (134600108 to W.L.).

Authors' contributions

WL, LS contributed to conception and design of the study and revised the manuscript. LS, WL, JD and YJ performed experiments and were responsible for data collection, analysis, and interpretation of the results. LW, NH and LL were responsible for conducting the data analysis. LS and WL drafted the manuscript and figures, and YL interpreted the data and critically revised the manuscript. All authors read and approved the final manuscript.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Original ArticlesMetastasis-associated in colon cancer-1 upregulates vascular endothelial growth factor-C/D to promote lymphangiogenesis in human gastric cancer

Highlights

Abstract

Introduction

Section snippets

Study population and tissue samples

Expression of MACC1 was correlated with LVD in human gastric cancer tissues

Discussion

Conclusions

Financial support

Authors' contributions

Conflict of interest

J. Biol. Chem

Acad. Radiol

Blood

Hum. Pathol

Cancer Lett

Mol. Oncol

Surgery

Cancer Cell

Cancer Cell

Cancer Lett

Lymphangiogenesis in development and human disease

Nature

Tumor lymphangiogenesis and metastatic spread-new players begin to emerge

Int. J. Cancer

Different significance between intratumoral and peritumoral lymphatic vessel density in gastric cancer: a retrospective study of 123 cases

BMC Cancer

Inhibition of cyclooxygenase-2 suppresses lymph node metastasis via reduction of lymphangiogenesis

Cancer Res

MACC1, a newly identified key regulator of HGF-MET signaling, predicts colon cancer metastasis

Nat. Med

Metastasis-associated in colon cancer-1 upregulation predicts a poor prognosis of gastric cancer, and promotes tumor cell proliferation and invasion

Int. J. Cancer

MACC1 down-regulation inhibits proliferation and tumourigenicity of nasopharyngeal carcinoma cells through Akt/beta-catenin signaling pathway

PLoS ONE

MACC1 overexpression predicts a poor prognosis for non-small cell lung cancer

Med. Oncol

MACC1 supports human gastric cancer growth under metabolic stress by enhancing the Warburg effect

Oncogene

Expression of MACC1 and c-Met in human gastric cancer and its clinical significance

Cancer Cell Int

Original Articles
Metastasis-associated in colon cancer-1 upregulates vascular endothelial growth factor-C/D to promote lymphangiogenesis in human gastric cancer