Original ArticlePancreatic stellate cells reorganize matrix components and lead pancreatic cancer invasion via the function of Endo180
Introduction
Pancreatic cancer is one of the most lethal cancers, and its 5-year survival rate is <8% [1], largely owing to the difficulty in early detection, fast progression (e.g., local invasion into surrounding organs and distant metastasis), and the poor improvements in treatments. Radical resection is the only curative treatment. However, pancreatic cancer patients that undergo radical operation have a poor prognosis for distant metastasis and local recurrence. Therefore, it is necessary to elucidate the pathogenesis of pancreatic cancer and understand the mechanisms of invasion and metastasis to develop a new treatment for the improvement of such a poor prognosis.
Pancreatic cancer is characterized by excessive desmoplasia with an abundant extracellular matrix (ECM), which plays a crucial role in its aggressive behavior [2], and resistance to traditional therapies through tumor-stromal interactions [3]. The enriched desmoplastic microenvironment in the ECM is mainly produced by activated pancreatic stellate cells (PSCs). They are activated upon stimulation by various autocrine and paracrine factors. PSCs express α-smooth muscle actin (αSMA) and produce various ECM proteins [4], [5]. PSC-derived soluble factors and an abundant ECM promote the proliferation, migration, and invasion of pancreatic cancer cells (PCCs), leading to poor survival of pancreatic cancer patients [6], [7].
During cancer cell invasion, the specific cell populations leading the local invasion are known as “leading cells”. Previously, Gaggioli et al. [8] reported that cancer-associated fibroblasts lead and promote squamous cancer cell invasion through matrix metalloproteinases (MMPs) and force-dependent generation of ECM tracks. Cheung et al. [9] reported that K14-positive cells directly modify tensile strength and viscoelastic properties to regulate cell–cell adhesion or change signal transduction mechanisms independently of their mechanical properties, leading to collective invasion of human breast cancer. Brentnall [10] reported that palladin-activated fibroblasts promote PCC invasion by creating tunnels though the ECM. However, the underlying mechanisms of the phenomenon have been unclear.
Remodeling of the ECM is considered to play a major role in pancreatic cancer invasion and metastasis [11]. ECM remodeling continues in cancer tissue and causes resistance to established therapies [12], [13]. Consequently, we need to understand the mechanisms of ECM remodeling to elucidate the invasive and metastatic behaviors of PCCs.
Endo180 (also known as CD280, MRC2, and urokinase-type plasminogen activator receptor-associated protein) is a multiple C-type lectin-like domain receptor that binds to extracellular collagens via its fibronectin type II domain [14]. It is essential for cellular uptake of collagen in fibroblasts and plays a physiological role in mediating collagen matrix remodeling during tissue development [14]. Furthermore, Endo180 induces Rho-ROCK-myosin light chain 2 (MLC2)-based contractile signals and promotes adhesion disassembly [15]. Endo180 is highly expressed in fibroblasts. Recently, Endo180 was found to be expressed in not only fibroblasts but also several cancers [16], [17], [18], [19]. Moreover, Endo180 is related to cancer cell survival in prostate cancer [18] and breast cancer [20]. These data suggest that Endo180 plays an important role in ECM remodeling and cancer progression. However, the mechanisms by which Endo180 contributes to the ECM remodeling of pancreatic cancer are not understood.
In the present study, we used in vitro three-dimensional (3D) matrix remodeling assay to identify the leading cells in pancreatic cancer and determine how these cells lead and promote cancer cell invasion in the ECM.
Section snippets
Materials and methods
Detailed information is provided in the Supplementary Materials and methods.
PSCs lead and promote local invasion of PCCs
To establish an in vitro model similar to the in vivo microenvironment of pancreatic cancer, we performed a 3D matrix remodeling assay in which PCCs and/or PSCs were embedded in the center of the collagen matrix (Fig. 1A). The movements of PSCs and PCCs were investigated in the surrounding collagen matrix, and we evaluated the effect of the physical behavior of PSCs on the invasive behavior of PCCs in the collagen matrix.
In the 3D matrix remodeling assay, PSC1 and SUIT2 more frequently invaded
Discussions
An abundant ECM is a major characteristic of pancreatic cancer. To reproduce the ECM, we used a 3D matrix remodeling assay and found that PSCs invaded into the collagen matrix and cancer cells frequently invaded following the invading PSCs in co-cultures of PCCs and PSCs. These observations suggest that PCCs and PSCs enhance the invasive abilities of each other by tumor-stromal (cancer cell-PSC) interactions as reported previously [2], [3], [23], [36], [37]. Furthermore, invading cancer cells
Acknowledgements
The authors thank E. Manbe, S. Sadatomi (Department of Surgery and Oncology, Kyushu University Hospital), and members of the Research Support Center and Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, for their expert technical assistance.
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