Stromal Fibroblasts Mediate Extracellular Matrix Remodeling and Invasion of Scirrhous Gastric Carcinoma Cells

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From: PLoS ONE(Vol. 9, Issue 1)
Publisher: Public Library of Science
Document Type: Article
Length: 7,105 words
Lexile Measure: 1370L

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Author(s): Hideki Yamaguchi 1, Nachi Yoshida 1, Miho Takanashi 1,2, Yuumi Ito 1,2, Kiyoko Fukami 2, Kazuyoshi Yanagihara 3, Masakazu Yashiro 4, Ryuichi Sakai 1,*


Recent studies have established the importance of the tumor stroma in cancer progression [1], [2]. Tumor stroma consists of many types of non-cancerous cells and non-cellular components including the extracellular matrix (ECM). Stromal fibroblasts (SFs) are major cellular constituents of tumor stroma and often called cancer-associated fibroblasts (CAFs) [3]. They have been implicated in the malignant behavior of cancers, such as cell proliferation, ECM remodeling, and angiogenesis [4]. Moreover, they often display the phenotypes of myofibroblasts, characterized by the expression of [alpha]-smooth muscle actin ([alpha]SMA) and strong contractility [5]. These characteristics contribute not only to fibrosis in tumor tissue but also to the remodeling and stiffening of the stromal ECM that are favorable for invasion and metastasis of carcinoma cells [6], [7].

Scirrhous gastric carcinoma (SGC), also known as diffusely infiltrative carcinoma, has a very poor prognosis due to rapid infiltrative invasion and a high incidence of peritoneal dissemination [8], [9]. SGC is associated with extensive stromal fibrosis, resulting in the thickening and hardening of the gastric wall and shrinkage of the stomach. As there is elevated proliferation of SFs in SGC lesions, they have been proposed to support the progression of this disease [10]. In fact, a positive correlation between the presence of SFs and the metastatic potential of gastric cancers has been found [11]. SGC cells induce fibrosis of the peritoneum in peritoneal dissemination, indicating that SFs also play a role here [12]. Recent studies have demonstrated that SFs stimulate migration and invasion of SGC cells [13], [14] and SGC cells reciprocally promote proliferation of gastric fibroblasts [14], [15]. However, the biological and molecular basis and the pathological function of the intercellular interaction between SGC cells and SFs remain largely unknown. In this study, we established the system to visualize and quantify the crosstalk between SFs and SGC cells for achieving the invasive properties and investigated the role of SFs in the invasion and remodeling of the ECM by SGC cells.

Materials and Methods

Cell culture

Human gastric cancer cell lines 58As9, HSC-59, HSC-44PE, and 44As3 were described previously [16], [17], and MKN1, MKN7, and MKN74 were obtained from the Health Science Research Resources Bank. 44As3 cells stably expressing tdTomato were generated by retroviral transduction. These cells were maintained in RPMI 1640 medium (Invitrogen) supplemented with 10% FBS and antibiotics at 37°C in a humidified atmosphere containing 5% CO2 . The orthotopic fibroblast cell lines, CaF37 and CaF38 were established from the tumoral gastric wall of SGC patients who had undergone gastrectomy. The primary gastric tumor was excised under aseptic conditions and minced with forceps and scissors. The tumor pieces were cultivated in DMEM (Nikken) with 10% FBS. After approximately 2 weeks, fibroblasts were collected and transferred to another culture dish. Serial passages were carried out every 4-7 days. The fibroblasts used were 4-10th passage of culture. This study was approved by the...

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Gale Document Number: GALE|A478870890