Elsevier

Journal of Proteomics

Volume 253, 20 February 2022, 104454
Journal of Proteomics

Matrisome provides a supportive microenvironment for oral squamous cell carcinoma progression

https://doi.org/10.1016/j.jprot.2021.104454Get rights and content

Highlights

  • An effective tissue engineering method is applied for remodeling ECM in vitro.

  • Proteomics analysis identifies differentially expressed proteins in ECM of oral squamous cell carcinoma and normal tissues.

  • The transformation of ECM has significant effects on OSCC progression.

Abstract

Oral squamous cell carcinoma (OSCC) is a common pernicious tumor in the head and neck regions. However, the function of tumor extracellular matrix (ECM) has not been elucidated. A tissue engineering method was applied for remodeling ECM through decellularization. The cellular components were removed, and the biological composition was mostly preserved. Proteomics was performed to analyze the characterization between normal and tumor ECM. According to LC-MS/MS results, 26 proteins just showed in tumor ECM, and 14 proteins only showed in late-stage tumor ECM. KEGG pathway analysis showed that most variant proteins were linked to metabolic regulation and tumor immunity (such as SCC-Ag1, LOX). To affirm the influence of tumor ECM on the progression of OSCC, tumor cells and macrophages were co-cultured with ECM scaffold. Marked differences in proliferation, apoptosis, and migration of OSCC cells were observed between tumor and normal ECM. Tumor ECM polarized macrophages towards an anti-inflammatory phenotype (higher IL-10 and CD68, and relatively lower CD86 and IL1-β). Collectively, these findings suggest that tumor ECM served as a permissive role in OSCC progression.

Significance

The variation between OSCC ECM and normal ECM confirm tumor ECM plays a significant role in OSCC deterioration, which is conducive to exploring the occurrence and progression mechanisms of OSCC, and further improving the curative effect of this disease.

Introduction

Oral tumor has become a significant health issue in recent years. Statistics indicate that the incidence of oral tumor ranks sixth among systemic malignancies worldwide, of which about 90% are squamous cell carcinoma. Although significant advancement has been witnessed in radical surgery in combination with auxiliary therapeutics (such as radiotherapy and chemotherapy), the five-year survival rate for patients has not prominently increased in recent decades [1]. Therefore, this calls for further studies to elucidate the underlying mechanism of OSCC progression in order to improve the available treatment strategies [2].

OSCC studies conducted in recent years have demonstrated that growth and aggression of the tumor are affected by the microenvironment around the tumor lesions [3,4]. It is well known that the inherent tumor microenvironment consists of several important portions, including ECM, cell–cell interaction, and cell–matrix communication [5,6]. Notably, ECM is a main constituent of the tumor microenvironment, which is composed of several networks of different biochemical components, including fibrins, glycoproteins, and polysaccharides [7]. Moreover, ECM plays an important role in morphogenesis, development, homoeostasis and disease through regulating cell-cell, cell- mesenchyme interaction [[8], [9], [10], [11]].

In recent years, decellularized ECM scaffold (dECM) from animal tissues has been developed as a prospective media for remodeling ECM in vitro [12,13]. The ECM scaffold almost completely gets rid of cellular components, and preserves the original ECM constitution and construction. The advent of proteomics has revolutionized studies on the composition of the ECM [14], and the term “Matrisome” is coined to define the protein composition of the ECM in the field of proteomics [15,16]. In this study, proteomics analysis was incorporated to determine the differences between normal and tumor ECM. These specific matrisome proteins might provide a new possible target for the treatment of this disease. To further confirm the role of ECM in OSCC progression, tumor cells and macrophages were co-cultured with ECM scaffold. Results showed that tumor ECM scaffolds maintained the biological support of OSCC cells for migration, invasion, and proliferation. They also distinctly modulated macrophages polarization activity.

The bioactive scaffold system developed in this study, through the decellularized approach, provides a new insight into the interactions between ECM environment and cells. The difference in tumor and normal oral ECM by mass spectroscopy contributes to unravelling the fundamental mechanisms of OSCC deterioration or therapeutic resistance.

Section snippets

Ethics and clinical samples

In this study, human OSCC and normal tissues which were adjacent to primary tumors were obtained after surgical operation in the Department of Oral & Maxillofacial Surgery, Stomatological Hospital of Chongqing Medical University. In this experiment, a total of 53 patients' samples were collected, these samples were staged according to the guidelines of AJCC8. Patients with tumor recurrence, oral metastasis of systemic tumors, history of radiotherapy and chemotherapy, HPV infection and other

Confirmation of decellularization efficacy

After decellularization, native normal and tumor tissues presented a striking change, not only in terms of color change (from pink to transparent white), but also the appearance (from solid to a gelatinous shape) (Fig. 1A). Histological examination, including H&E and Masson's Trichrome staining, affirmed an absolute elimination of nuclear staining, and showed a random distribution of muscle and collagen fibers in the decellularized tissues (Fig. 1B–C). To further evaluate the efficiency of

Discussion

Studies have shown that tumors originate from genetic mutation, which causes uncontrolled cell biological activities by altering cellular functions such as survival and apoptosis [25]. In recent years, limited breakthroughs have been made in OSCC fundamental research [2]. As an important part of tumor microenvironment, ECM not only serves as a superior biological scaffold structure, but also regulates cell growth, metabolism, and differentiation through various signaling molecules [29,32].

In

Conclusion

This study shows that detergent immersion methods can effectively prepare ECM scaffold from both human oral tissues. Based on the results of the proteomics analysis, the research demonstrates that tumor ECM is reconstructed during OSCC progression. The transformation of ECM has significant effects on the behavior of tumor cells, immune cells, and other cells. The research is still in its initial stage, further work needs to elucidate the mechanism of ECM proteins related to OSCC progression,

Declaration of Competing Interest

The authors declare no conflict of interest.

Acknowledgments

This study was funded by Natural Science Foundation of Chongqing (Grant No. cstc2019jscx-msxm X0173) and Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant No. KJQN202000417). We thank the Shanghai Applied Protein Technology Co., Ltd., for technological assistance.

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