TIMP-2 secreted by monocyte-like cells is a potent suppressor of invadopodia formation in pancreatic cancer cells

Christian Benzing,Chi Man Tsang,Alexander Rimmer,Hoyin Lam,Yoana Arroyo-Berdugo,Yolanda Calle,Claire M. Wells

doi:10.1186/s12885-019-6429-z

Abstract

BackgroundMonocytes are a major component of the tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC). However, the complex interactions between tumor cells and monocytes and their role in tumor invasion have not been fully established.MethodsTo specifically test the impact of interaction on invasive potential two PDAC cell lines PaTu8902 and CFPAC-1 were selected on their ability to form invasive adhesions, otherwise known as invadopodia and invade in a spheroid invasion assay.ResultsInterestingly when the PDAC cells were co-cultured with undifferentiated THP1 monocyte-like cells invadopodia formation was significantly suppressed. Moreover, conditioned media of THP1 cells (CM) was also able to suppress invadopodia formation. Further investigation revealed that both tissue inhibitor of metalloproteinase (TIMP) 1 and 2 were present in the CM. However, suppression of invadopodia formation was found that was specific to TIMP2 activity.ConclusionsOur findings indicate that TIMP2 levels in the tumour microenvironment may have prognostic value in patients with PDAC. Furthermore, activation of TIMP2 expressing monocytes in the primary tumour could present a potential therapeutic opportunity to suppress cell invasion in PDAC.

Highlights

Monocytes are a major component of the tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC)
Growth medium in plate 2 and 3 were removed from the wells and replaced with a) normal growth medium (DMEM supplemented with 10% fetal bovine serum (FBS) and 1 mM penicillin/streptomycin), b) Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS and 1 mM penicillin/streptomycin mixed with serum-free DMEM (1:1), c) (DMEM supplemented with 10% FBS and 1 mM penicillin/streptomycin mixed with frozen and rewarmed THP1-conditioned media (CM) (1:1), d) DMEM supplemented with 10% FBS and 1 mM penicillin/ streptomycin containing recombinant TIMP2 (rTIMP2) (50 ng/ml)
We found that Pancreatic ductal adenocarcinoma (PDAC) cells require longer incubation times than previously reported for breast cancer cells [23] to generate quantifiable degradation activity and quantification of activity was based on the total area of gelatin degradation per field of view

Summary

Introduction

Monocytes are a major component of the tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC). The complex interactions between tumor cells and monocytes and their role in tumor invasion have not been fully established. Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease characterised by an aggressive biological tumour behaviour leading to a high mortality rate [1]. The invasive character of PDACs leads to a rapid progression of the disease [2]. In order to invade the surrounding tissue or to create distant metastases, cancer cells need to break through the basement membrane and degrade the extracellular matrix. One identified strategy is to utilise actin-rich membrane protrusions called invadopodia [3] that can degrade extracellular matrix. Initiating mechanisms to suppress such formation is a potential therapeutic target [4].

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