The composition of prostate core matrisome in vivo and in vitro unveiled by mass spectrometric analysis

Ojalill, Marjaana; Rappu, Pekka; Siljamäki, Elina; Taimen, Pekka; Boström, Peter J.; Heino, Jyrki

doi:10.1002/pros.23503

Cited by 11 publications

(11 citation statements)

References 51 publications

(98 reference statements)

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“…None of the common or DU145‐specific gene products represented core matrisome structural proteins, while PC3 cells could produce laminin chains α3, β3, γ2, as well as HSPG2 and collagen VI (COL6A1) (Figure 1B; Table SI). In accordance with our previous observations, 21 fibroblastic cells were responsible for the production of the structural proteins in the core matrisome (Figure 1C; Table 1). The major components of collagenous, fibronectin‐rich ECM could be detected in fibroblast spheroids, including the two α chains of fibril forming collagen I (COL1A1 and COL1A2), fibril‐associated proteins, such as collagen XII (COL12A1) and decorin (DCN), tenascin C (TNC), and beaded filament‐forming collagen VI (COL6A2 and COL6A3) (Table 1).…”

Section: Resultssupporting

confidence: 93%

“…The human androgen‐independent prostate epithelial adenocarcinoma cell lines DU145 and PC3 (both from American Type Culture Collection) and human prostate‐derived fibroblastic cells, described previously 21 were cultured in Roswell Park Memorial Institute (RPMI)‐1640 medium (Lonza), supplemented with 10% fetal calf serum (FCS, Biowest), 2 mM Ultraglutamate, 100 U/mL penicillin and streptomycin (Lonza). In the indicated experiments also serum‐free (SF) RPMI‐1640 medium or keratinocyte serum‐free medium (KSFM) with 5 ng/mL human recombinant epidermal growth factor, 50 μg/mL bovine pituitary extract (Gibco), supplemented with 2 ng/mL recombinant human leukemia inhibitory factor and 2 ng/mL stem cell factor (Sigma Aldrich) was used.…”

Section: Methodsmentioning

confidence: 99%

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Interaction between prostate cancer cells and prostate fibroblasts promotes accumulation and proteolytic processing of basement membrane proteins

et al. 2020

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Background: Tumor microenvironment or stroma has the potency to regulate the behavior of malignant cells. Fibroblast-like cells are abundant in tumor stroma and they are also responsible for the synthesis of many extracellular matrix components.Fibroblast-cancer cell interplay can modify the functions of both cell types. Methods:We applied mass spectrometry and proteomics to unveil the matrisome in 3D spheroids formed by DU145 prostate cancer cells, PC3 prostate cancer cells, or prostate-derived fibroblasts. Similarly, DU145/fibroblast and PC3/fibroblast coculture spheroids were also analyzed. Western blot analysis and immunofluorescence were used to confirm the presence of specific proteins in spheroids. Cancer dissemination was studied by utilizing "out of spheroids" migration and invasion assays.Results: In the spheroid model cancer cell-fibroblast interplay caused remarkable changes in the extracellular matrix and accelerated the invasion of DU145 cells.Fibroblasts produced structural matrix proteins, growth factors, and matrix metalloproteinases. In cancer cell/fibroblast cocultures basement membrane components, including laminins (α3, α5, β2, and β3), heparan sulfate proteoglycan (HSPG2 gene product), and collagen XVIII accumulated in a prominent manner when compared with spheroids that contained fibroblasts or cancer cells only. Furthermore, collagen XVIII was intensively processed to different endostatin-containing isoforms by cancer cell-derived cathepsin L. Conclusions: Fibroblasts can promote carcinoma cell dissemination by several different mechanisms. Extracellular matrix and basement membrane proteins provide attachment sites for cell locomotion promoting adhesion receptors. Growth factors and metalloproteinases are known to accelerate cell invasion. In addition, cancer cell-fibroblast interplay generates biologically active fragments of basement membrane proteins, such as endostatin.

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Section: Resultssupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Interaction between prostate cancer cells and prostate fibroblasts promotes accumulation and proteolytic processing of basement membrane proteins

et al. 2020

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“…ECM-enriched protein extracts from tumorsphere cultures in PromoCell were prepared by trypsin digestion as described (Ojalill et al, 2018). Peptides were separated by a nanoflow HPLC system (Easy-nLC1000, Thermo) coupled to a Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer (Thermo).…”

Section: Methodsmentioning

confidence: 99%

FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy

Osterman¹,

Ozmadenci²,

Kleinschmidt³

et al. 2019

eLife

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Gene copy number alterations, tumor cell stemness, and the development of platinum chemotherapy resistance contribute to high-grade serous ovarian cancer (HGSOC) recurrence. Stem phenotypes involving Wnt-β-catenin, aldehyde dehydrogenase activities, intrinsic platinum resistance, and tumorsphere formation are here associated with spontaneous gains in Kras, Myc and FAK (KMF) genes in a new aggressive murine model of ovarian cancer. Adhesion-independent FAK signaling sustained KMF and human tumorsphere proliferation as well as resistance to cisplatin cytotoxicity. Platinum-resistant tumorspheres can acquire a dependence on FAK for growth. Accordingly, increased FAK tyrosine phosphorylation was observed within HGSOC patient tumors surviving neo-adjuvant chemotherapy. Combining a FAK inhibitor with platinum overcame chemoresistance and triggered cell apoptosis. FAK transcriptomic analyses across knockout and reconstituted cells identified 135 targets, elevated in HGSOC, that were regulated by FAK activity and β-catenin including Myc, pluripotency and DNA repair genes. These studies reveal an oncogenic FAK signaling role supporting chemoresistance.

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“…Highly controlled and reproducible renal TME created with collagen-based bioink has shown potential applications for anticancer drug screening [ 86 ]. In prostate, collagen comprises more than 12% among the 120 types of ECM proteins [ 87 , 88 ], and the deregulation of collagen metabolism was also correlated with PCa Gleason score and could be a predictor of patient survival [ [89] , [90] , [91] , [92] ]. In bladder, collagen is commonly found in the connective tissue outside the muscle bundles of bladder wall [ 93 , 94 ] and the ratio of type III to type I collagen has been observed to be abnormally elevated in patients with bladder disease compared with normal bladder [ [95] , [96] , [97] ].…”

Section: Commonly Used 3d Bioprinting Technologies In Urological Dise...mentioning

confidence: 99%

The application of 3D bioprinting in urological diseases

Han

Huang

et al. 2022

Materials Today Bio

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The composition of prostate core matrisome in vivo and in vitro unveiled by mass spectrometric analysis

Abstract: Artificial activation of ECM remodeling seems to be a distinctive feature in in vitro models. In conclusion the constitution of ECM produced by prostate derived fibroblasts in vitro is similar, but not identical to the prostate ECM in vivo as shown here by mass spectrometric analysis.

Cited by 11 publications

References 51 publications

Interaction between prostate cancer cells and prostate fibroblasts promotes accumulation and proteolytic processing of basement membrane proteins

Interaction between prostate cancer cells and prostate fibroblasts promotes accumulation and proteolytic processing of basement membrane proteins

FAK activity sustains intrinsic and acquired ovarian cancer resistance to platinum chemotherapy

The application of 3D bioprinting in urological diseases

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