Pancreatic Stellate Cells Potentiate Proinvasive Effects of SERPINE2 Expression in Pancreatic Cancer Xenograft Tumors

doi:10.1159/000107400

Pancreatology

Volume 7, Issue 4, September 2007, Pages 380-385

https://doi.org/10.1159/000107400 Get rights and content

Abstract

We have previously reported that inducible overexpression of the serine protease inhibitor nexin 2 (SERPINE2) significantly increases local invasiveness of subclones of the pancreatic cancer cell-line SUIT-2 in nude mouse xenografts. This was associated with a striking increase of extracellular matrix deposition in the invasive tumors. Pancreatic stellate cells (PSCs) have recently been identified as the major source of fibrosis in pancreatic adenocarcinomas. Here we report that co-injection of PSCs and tumor cells dramatically enhances the invasive potential of serine protease inhibitor Nexin 2 (SERPINE2)-expressing SUIT-2 cells. 100% (24 of 24) of the SERPINE2-expressing tumors with PSCs grew aggressively invasive, as compared to 39% of SERPINE2-negative tumors with PSCs and 27% of SERPINE2-expressing tumors without PSCs. In contrast to pure cancer cell preparations, SERPINE2 overexpression in the presence of PSCs also resulted in increased tumor growth. Histological evaluation demonstrated the presence of large amounts of ECM deposits co-localizing with cells staining positive for the PSC marker α-SMA. We conclude that PSCs actively proliferate in pancreatic cancer xenograft tumors and significantly contribute to the local invasive potential of the tumors. Presence of PSCs enhances the pro-invasive effects of SERPINE2 expression, and SERPINE2 influences tumor growth (as opposed to invasiveness) only in the presence of PSCs. Our data thus suggest that SERPINE2 is an important modulator of tumor cell/host interactions in pancreatic cancer.

References (27)

MG Bachem et al.
Identification, culture, and characterization of pancreatic stellate cells in rats and humans
Gastroenterology
(1998)
MG Bachem et al.
Pancreatic carcinoma cells induce fibrosis by stimulating proliferation and matrix synthesis of stellate cells
Gastroenterology
(2005)
S Yoshida et al.
Pancreatic cancer stimulates pancreatic stellate cell proliferation and TIMP-1 production through the MAP kinase pathway
Biochem Biophys Res Commun
(2004)
JA Kiefer et al.
Type I collagen-mediated proliferation of PC3 prostate carcinoma cell line: implications for enhanced growth in the bone microenvironment
Matrix Biol
(2001)
R Paddenberg et al.
Phenotypical changes of a human pancreatic adenocarcinoma cell line after selection on laminin-1/nidogen (LM/Ng) substratum
Eur J Cell Biol
(1998)
CK Weber et al.
Biglycan is overexpressed in pancreatic cancer and induces G1-arrest in pancreatic cancer cell lines
Gastroenterology
(2001)
H Yamanari et al.
Extracellular matrix components regulating glandular differentiation and the formation of basal lamina of ahuman pancreatic cancer cell line in vitro
Exp Cell Res
(1994)
J Mollenhauer et al.
Distribution of extracellular matrix proteins in pancreatic ductal adenocarcinoma and its influence on tumor cell proliferation in vitro
Pancreas
(1987)
TM Gress et al.
Expression and in-situ localization of genes coding for extracellular matrix proteins and extracellular matrix degrading proteases in pancreatic cancer
Int J Cancer
(1995)
IJ Fidler
Critical factors in the biology of human cancer metastasis: twenty-eighth G.H.A
Clowes memorial award lecture. Cancer Res
(1990)

R Fridman et al.

Reconstituted basement membrane (matrigel) and laminin can enhance the tumorigenicity and the drug resistance of small cell lung cancer cell lines

Proc Natl Acad Sci USA

(1990)

S Muerkoster et al.

Tumor stroma interactions induce chemoresistance in pancreatic ductal carcinoma cells involving increased secretion and paracrine effects of nitric oxide and interleukin-1beta

Cancer Res

(2004)

T Sethi et al.

Extracellular matrix proteins protect small cell lung cancer cells against apoptosis: a mechanism for small cell lung cancer growth and drug resistance in vivo

Nat Med

(1999)

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Pancreatic stellate cells: Aiding and abetting pancreatic cancer progression
2020, Pancreatology
Citation Excerpt :
Tian and colleagues (2012) have reported that the binding of the fibroblast growth factor receptor (FGFR) expressed on PSCs to its ligand FGF secreted by cancer cells induces an invasive phenotype of pancreatic cancer. Studies on in vivo models have demonstrated the modulatory function by PSCs and their role in potentiating the invasiveness of PC cells expressing serine protease inhibitor nexin2 (SERPINE2) possibly by modulating the ECM production and matrix reorganization in the tumour [62]. Another growth factor pathway of interest in PC, is the HGF-c-MET pathway.
Tumour-stromal interactions have now been acknowledged to play a major role in pancreatic cancer (PC) progression. The abundant collagenous stroma is produced by a specific cell type in the pancreas-the pancreatic stellate cell (PSC). Pancreatic stellate cells (PSCs) are a unique resident cell type of pancreas and with a critical role in both healthy and diseased pancreas. Accumulating evidence indicates that PSCs interact closely with cancer cells as well as with other cell types of the stroma such as immune cells, endothelial cells and neuronal cells, to set up a growth permissive microenvironment for pancreatic tumours, which facilitates local tumour growth as well as distant metastasis. Consequently, recent work in the field has focused on the development of novel therapeutic approaches targeting the stroma to inhibit PC progression. Such a multi-pronged approach targeting both tumour and stromal elements of PC has been successfully applied in pre-clinical settings. The challenge now is to translate the pre-clinical findings into the clinical setting to achieve better outcomes for pancreatic cancer patients.
Mitogen Inducible Gene-6 Is a Prognostic Marker for Patients with Colorectal Liver Metastases
2019, Translational Oncology
Citation Excerpt :
We found 68 genes that were consistently up-regulated and 76 that were down-regulated in CRCLM relative to matched primary tumors. From this group of 144 differentially expressed genes, 7 were selected for validation based on the association between mRNA level (using values obtained from the microarray) and clinical outcome (Table 3): lumican, a proteoglycan of the extracellular matrix reported to be overexpressed in colorectal breast, neuroendocrine, and other cancers [21]; tissue inhibitor of metalloproteinase 1 (TIMP1), an inhibitor of matrix metalloproteinases and modulator of other diverse processes in cancer cells [22]; basic helix-loop-helix domain containing B2 (BHLHB2), a transcription factor that is induced by hypoxia within tumors [23]; fibronectin, an abundant extracellular matrix protein that contributes to altered stromal remodeling during tumorigenesis [24]; TM4SF1 (transmembrane 4 superfamily member 1), a tumor-associated antigen that regulates cancer cell motility and invasion [25]; serpine2, an extracellular serine protease that is overexpressed in pancreatic, colon, and stomach cancers that may facilitate invasive processes [26]; and MIG-6, a putative tumor suppressor that negatively regulates the ErbB family of receptor tyrosine kinases [27,28]. To determine whether the expression of the candidate genes identified above as correlating with clinical outcome for unresectable patients with colorectal metastases confined to the liver also applied to patients undergoing potentially curative hepatectomy for CRCLM, we performed qRT-PCR of the 7 candidate genes on CRCLM from 56 additional patients.
PURPOSE: Prognostic schemes that rely on clinical variables to predict outcome after resection of colorectal metastases remain imperfect. We hypothesized that molecular markers can improve the accuracy of prognostic schemes. METHODS: We screened the transcriptome of matched colorectal liver metastases (CRCLM) and primary tumors from 42 patients with unresected CRCLM to identify differentially expressed genes. Among the differentially expressed genes identified, we looked for associations between expression and time to disease progression or overall survival. To validate such associations, mRNA levels of the candidate genes were assayed by qRT-PCR from CRCLM in 56 additional patients who underwent hepatectomy. RESULTS: Seven candidate genes were selected for validation based on their differential expression between metastases and primary tumors and a correlation between expression and surgical outcome: lumican; tissue inhibitor metalloproteinase 1; basic helix-loop-helix domain containing class B2; fibronectin; transmembrane 4 superfamily member 1; mitogen inducible gene 6 (MIG-6); and serpine 2. In the hepatectomy group, only MIG-6 expression was predictive of poor survival after hepatectomy. Quantitative PCR of MIG-6 mRNA was performed on 25 additional hepatectomy patients to determine if MIG-6 expression could substratify patients beyond the clinical risk score. Patients within defined clinical risk score categories were effectively substratified into distinct groups by relative MIG-6 expression. CONCLUSIONS: MIG-6 expression is inversely associated with survival after hepatectomy and may be used to improve traditional prognostic schemes that rely on clinicopathologic data such as the Clinical Risk Score.
Matrix control of pancreatic cancer: New insights into fibronectin signaling
2016, Cancer Letters
Citation Excerpt :
Stromal cells found within the tumor microenvironment can promote PDA tumorigenicity and chemoresistance by enabling ECM–cancer cell interaction [62,63]. For example, subcutaneous co-injection of pancreatic cancer cells and PSCs resulted in larger tumors compared to those mice that were injected with cancer cells only [64,65]. These co-injected tumors showed an increased fibrotic response and increased cancer cell proliferation.
Pancreatic ductal adenocarcinoma (PDA) is a highly metastatic disease that resists most current therapies. A defining characteristic of PDA is an intense fibrotic response that promotes tumor cell invasion and chemoresistance. Efforts to understand the complex relationship between the tumor and its extracellular network and to therapeutically perturb tumor–stroma interactions are ongoing. Fibronectin (FN), a provisional matrix protein abundant in PDA stroma but not normal tissues, supports metastatic spread and chemoresistance of this deadly disease. FN also supports angiogenesis, which is required for even hypovascular tumors such as PDA to develop and progress. Targeting components of the tumor stroma, such as FN, can effectively reduce tumor growth and spread while also enhancing delivery of chemotherapy. Here, we review the molecular mechanisms by which FN drives angiogenesis, metastasis and chemoresistance in PDA. In light of these new findings, we also discuss therapeutic strategies to inhibit FN signaling.
Pancreatic stellate cells and pancreas cancer: Current perspectives and future strategies
2014, European Journal of Cancer
Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant disease with a very poor prognosis. To date patient outcomes have not improved principally due to the limited number of patients suitable for surgical resections and the radiation and chemotherapy resistance of these tumours. In the last decade, a failure of conventional therapies has forced researchers to re-examine the environment of PDAC. The tumour environment has been demonstrated to consist of an abundance of stroma containing many cells but predominantly pancreatic stellate cells (PSCs). Recent research has focused on understanding the interaction between PSCs and PDAC cells in vitro and in vivo. It is believed that the interaction between these cells is responsible for supporting tumour growth, invasion and metastasis and creating the barrier to delivery of chemotherapeutics. Novel approaches which focus on the interactions between PDAC and PSCs which sustain the tumour microenvironment may achieve significant patient benefits. This manuscript reviews the current evidence regarding PSCs, their interaction with PDAC cells and the potential implication this may have for future therapies.
A PubMed search was carried out for the terms ‘pancreas cancer’ OR ‘pancreatic cancer’, AND ‘pancreatic stellate cells’, NOT ‘hepatic stellate cells’. All studies were screened and assessed for their eligibility and manuscripts exploring the relationship between PSCs and PDAC were included. The studies were subdivided into in vitro and in vivo groups.
One hundred and sixty-six manuscripts were identified and reduced to seventy-three in vitro and in vivo studies for review. The manuscripts showed that PDAC cells and PSCs interact with each other to enhance proliferation, reduce apoptosis and increase migration and invasion of cancer cells. The pathways through which they facilitate these actions provide potential targets for future novel therapies.
There is accumulating evidence supporting the multiple roles of PSCs in establishing the tumour microenvironment and supporting the survival of PDAC. To further validate these findings there is a need for greater use of physiologically relevant models of pancreatic cancer in vitro such as three dimensional co-cultures and the use of orthotopic and genetically engineered murine (GEM) models in vivo.
The stromal compartments in pancreatic cancer: Are there any therapeutic targets?
2014, Cancer Letters
Citation Excerpt :
For example, the expression of EMMPRIN (extracellular matrix metalloproteinase inducer) by cancer cells induces the production of MMP-2 by PSCs resulting in increased invasion of the tumour cells [102]. Also, serine protease inhibitor nexin 1 (SERPINE2) expressed by tumour cells promotes invasion and tumour growth only in presence of PSCs [103]. It is suggested that SERPINE2 mediates this effect through the stimulation of ECM protein expression by PSCs [104].
Pancreatic ductal adenocarcinoma (PDAC) is characterised by an abundant stromal response also known as a desmoplastic reaction. Pancreatic Stellate Cells have been identified as playing a key role in pancreatic cancer desmoplasia. There is accumulating evidence that the stroma contributes to tumour progression and to the low therapeutic response of PDAC patients. In this review we described the main actors of the desmoplastic reaction within PDAC and novel therapeutic approaches that are being tested to block the detrimental function of the stroma.
Pancreatic cancer: The role of pancreatic stellate cells in tumor progression
2011, Pancreatology
Pancreatic ductal adenocarcinoma is an aggressive and highly lethal disease frequently characterized by a dense stromal or desmoplastic response. Accumulating evidence exists that tumor desmoplasia plays a central role in disease progression and that e.g. activated pancreatic stellate cells (PSCs) are responsible for the excess matrix production. The mechanisms underlying the tumor versus stroma interplay are complex. Pancreatic cancer cells release mitogenic and fibrogenic stimulants, such as transforming growth factor β₁ platelet-derived growth factor (PDGF), sonic hedgehog, galectin 3, endothelin 1 and serine protease inhibitor nexin 2, all of which may promote the activated PSC phenotype. Stellate cells in turn secrete various factors, including PDGF, stromal-derived factor 1, epidermal growth factor, insulin-like growth factor 1, fibroblast growth factor, secreted protein acidic and rich in cysteine, matrix metalloproteinases, small leucine-rich proteoglycans, periostin and collagen type I that mediate effects on tumor growth, invasion, metastasis and resistance to chemotherapy. This review intends to shed light on the mechanisms by which PSCs in the stroma influence pancreatic cancer development. The increased understanding of this interaction will be of potential value in designing new modalities of targeted therapy.

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¹: Prof. Dr. Thomas M. Gress Division of Gastroenterology and Endocrinology University Hospital Giessen and Marburg, site Marburg Philipps University Marburg, Baldinger Strasse, DE-35043 Marburg (Germany) Tel. +49 6421 28 66 460, Fax +49 6421 28 68 922

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Pancreatic Stellate Cells Potentiate Proinvasive Effects of SERPINE2 Expression in Pancreatic Cancer Xenograft Tumors

Abstract

Gastroenterology

Gastroenterology

Biochem Biophys Res Commun

Matrix Biol

Eur J Cell Biol

Gastroenterology

Exp Cell Res

Distribution of extracellular matrix proteins in pancreatic ductal adenocarcinoma and its influence on tumor cell proliferation in vitro

Pancreas

Expression and in-situ localization of genes coding for extracellular matrix proteins and extracellular matrix degrading proteases in pancreatic cancer

Int J Cancer

Critical factors in the biology of human cancer metastasis: twenty-eighth G.H.A

Clowes memorial award lecture. Cancer Res

Reconstituted basement membrane (matrigel) and laminin can enhance the tumorigenicity and the drug resistance of small cell lung cancer cell lines

Proc Natl Acad Sci USA

Tumor stroma interactions induce chemoresistance in pancreatic ductal carcinoma cells involving increased secretion and paracrine effects of nitric oxide and interleukin-1beta

Cancer Res

Extracellular matrix proteins protect small cell lung cancer cells against apoptosis: a mechanism for small cell lung cancer growth and drug resistance in vivo

Nat Med