Proliferation Of Stromal Fibroblasts Research Articles

Molecular and functional characterization of tumor-induced factor (TIF): Hamster homolog of CXCL3 (GROγ) displays tumor suppressive activity

Previously our lab has created a mouse ovarian xenograft model of copy number variation (CNV)-mediated G protein-coupled receptor (GPCR) MAS-driven tumorigenesis, and RNA profiling identified a putative chemokine tumor-induced factor (Tif). Sequence analysis and chemotactic study suggested that Tif was likely to be a hamster homolog of human GROγ (CXCL3) [IJC 125 (2009): 1316–1327]. In the present study, we report the molecular and functional characterization of the Tif gene. Genomic study of CHO-K1 cells indicated that Tif gene consisted of 4 exons, characterized with an antisense B1 element which is embedded in the fourth exon. Two Tif transcripts were identified which shared identical sequences except that a string of 71-nt derived from the antisense B1 element was deficient in the shorter transcript. Of interests, B1-like RNA ladder was detected in xenografts. Functional studies showed that TIF induced chemotaxis and neovessel formation. Pharmacological studies suggested that TIF activated Gi-coupled CXCR2 and induced both calcium mobilization and ERK1/2 phosphorylation, and suppressed forskolin-stimulated cAMP accumulation. In addition, secreted matured TIF functioned as an autocrine factor and promoted anchorage-independent growth. Unexpectedly, TIF delayed the onset of tumor formation, possibly via suppressing proliferation of stromal fibroblasts. However, TIF did not exert any inhibitory effect on tumor growth. Potentially, TIF could be used for preventing cancer relapse.

Pancreatic cancer: Current status and Challenges.

The 5-year survival rate of patients with pancreatic cancer (PanCA) has remained stagnant. Unfortunately, the incidence is almost equal to mortality rates. These facts underscore the importance of concerted efforts to understand the pathology of this disease. Deregulation of multiple signaling pathways involved in a wide variety of cellular processes including proliferation, apoptosis, invasion, and metastasis contribute not only to cancer development but also to therapeutic resistance. The purpose of this review is to summarize current understanding of etiological factors including emerging evidence on the role of infectious agents, factors associated with therapeutic resistance and therapeutic options. The unique aspect of PanCA is "desmoplasia", a process that involves proliferation of stromal fibroblasts and collagen deposition in and around the filtrating cancer. Recent studies have identified pancreatic stellate cells (PSCs) as a potential source of such desmoplasia. Biphasic interactions between PSCs and cancer cells, endothelial cells, and/or myeloid derived suppressor cells in the tumor microenvironment contribute to pancreatic carcinogenesis. We summarize limitations of current therapeutic approaches and potential strategies to overcome these limitations using natural products including botanicals as adjuvant/neo-adjuvant for effective management of PanCA.

Inflammatory responses of stromal fibroblasts to inflammatory epithelial cells are involved in the pathogenesis of bovine mastitis

Hypernomic secretion of epithelial cytokines has several effects on stromal cells. The contributions of inflammatory epithelial cells to stromal fibroblasts in bovine mammary glands with mastitis remain poorly understood. Here, we established an inflammatory epithelial cell model of bovine mastitis with gram-negative lipopolysaccharide (LPS) and gram-positive lipoteichoic acid (LTA) bacterial cell wall components. We characterized immune responses of mammary stromal fibroblasts induced by inflammatory epithelial cells. Our results showed that inflammatory epithelial cells affected stromal fibroblast characteristics by increasing inflammatory mediator expression, elevating extracellular matrix protein deposition, decreasing proliferation capacity, and enhancing migration ability. The changes in stromal fibroblast proliferation and migration abilities were mediated by signal molecules, such as WNT signal pathway components. LPS- and LTA-induced inflammatory epithelial cells triggered different immune responses in stromal fibroblasts. Thus, in mastitis, bovine mammary gland stromal fibroblasts were affected by inflammatory epithelial cells and displayed inflammation-specific changes, suggesting that fibroblasts play crucial roles in bovine mastitis.

A Rare Mimicker in the Placenta.

A 24-year-old primigravida with monochorionic monoamniotic pregnancy presented to our institution at 31+3 weeks of gestation with leaking per vaginum. The ultrasound findings were that of an enlarged and cystically dilated placenta, which raised the possibility of a differential diagnosis of partial mole. In view of foetal distress she was taken up for emergency caesarean section. Two healthy female twins were delivered with no features of Beckwith Wiedermann syndrome. On gross examination the placenta weighed 800 gm and measured 17x15x2.5cm. The two umbilical cords attached measured 20cm and 24cm respectively [Table/Fig-1]. Foetal surface showed prominent, dilated blood vessels. The maternal surface was intact with cotyledons. Tiny, white nodules were seen on the foetal side. The cut surface of the placenta was beefy red with no pale areas. A focal area showed pinkish white vesicle like area [Table/Fig-2]. [Table/Fig-1]: Enlarged placenta (M) 17x15x2.5cm with two umbilical cords. [Table/Fig-2]: Placenta showing multiple grape like vesicles. The umbilical cords showed three vessels each. Multiple sections from the placenta showed villi of varying sizes [Table/Fig-3]. Some of the stem villi appeared markedly hydropic with stromal fibroblast proliferation. Concentric fibrosis was seen around the blood vessels [Table/Fig-4]. There was thick walled blood vessels with organising thrombus formation noted [Table/Fig-5]. Rest of the villi showed chorangiosis like change [Table/Fig-6]. Few villi showed central cistern formation, however, there was no trophoblastic proliferation. Sections from the villi appeared unremarkable. [Table/Fig-3]: Histopathology showed dilated stem villi (H&E 40x). [Table/Fig-4]: Histopathology showed concentric fibrosis in the villi (H&E 100x). [Table/Fig-5]: Histopathology showed organising thrombus (H & E 100x). [Table/Fig-6]: Histopathology showed increased number of blood vessels per villi, showing chorangiosis like change (H&E 200x).

Abstract 3601: Stromal fibroblasts mediate extracellular matrix remodeling and invasion of scirrhous gastric carcinoma cells

Abstract Scirrhous gastric carcinoma (SGC; also called diffusely infiltrative carcinoma, Borrmann's type-IV carcinoma, or the linitis plastica-type carcinoma) has the worst prognosis of all gastric cancers, owing to its rapid expansion by invasion and frequent peritoneal dissemination. Due to the increased proliferation of stromal fibroblasts (SFs) that occurs within SGC lesions and the peritoneal metastatic sites, SFs have been proposed to support the progression of this disease. However, the biological and molecular basis and the pathological role of the intercellular interaction between SGC cells and SFs remain largely unknown. In this study, we investigated the role of SFs in the invasion of the extracellular matrix (ECM) by SGC cells. When SGC cells were cocultured with SFs derived from SGC tissue on three-dimensional (3D) Matrigel, they were attracted together to form large cellular aggregates that invaded within the Matrigel. Time-lapse imaging revealed that this process was associated with extensive contraction and remodeling of the ECM. Immunofluorescence and biochemical analysis showed that SGC cells stimulate phosphorylation of myosin light chain and actomyosin-mediated mechanical remodeling of the ECM by SFs. By utilizing this assay system for inhibitor library screening, we have identified several inhibitors that potently suppress the cooperation between SGC cells and SFs to form the invasive structures. Among them, a Src inhibitor dasatinib impaired the interaction between SGC cells and SFs both in vitro and in vivo and effectively blocked peritoneal dissemination of SGC cells. These results indicate that SFs mediate mechanical remodeling of the ECM by SGC cells, thereby promoting invasion and peritoneal dissemination of SGC. Citation Format: Hideki Yamaguchi, Kazuyoshi Yanagihara, Masakazu Yashiro, Ryuichi Sakai. Stromal fibroblasts mediate extracellular matrix remodeling and invasion of scirrhous gastric carcinoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3601. doi:10.1158/1538-7445.AM2014-3601

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

Scirrhous gastric carcinoma (SGC) has the worst prognosis of all gastric cancers, owing to its rapid expansion by invasion and frequent peritoneal dissemination. Due to the increased proliferation of stromal fibroblasts (SFs) that occurs within SGC lesions and the peritoneal metastatic sites, SFs have been proposed to support the progression of this disease. However, the biological and molecular basis and the pathological role of the intercellular interaction between SGC cells and SFs remain largely unknown. In this study, we investigated the role of SFs in the invasion of the extracellular matrix (ECM) by SGC cells. When SGC cells were cocultured with SFs derived from SGC tissue on three-dimensional (3D) Matrigel, they were attracted together to form large cellular aggregates that invaded within the Matrigel. Time-lapse imaging revealed that this process was associated with extensive contraction and remodeling of the ECM. Immunofluorescence and biochemical analysis showed that SGC cells stimulate phosphorylation of myosin light chain and actomyosin-mediated mechanical remodeling of the ECM by SFs. By utilizing this assay system for inhibitor library screening, we have identified several inhibitors that potently suppress the cooperation between SGC cells and SFs to form the invasive structures. Among them, a Src inhibitor dasatinib impaired the interaction between SGC cells and SFs both in vitro and in vivo and effectively blocked peritoneal dissemination of SGC cells. These results indicate that SFs mediate mechanical remodeling of the ECM by SGC cells, thereby promoting invasion and peritoneal dissemination of SGC.

Abstract 47: Cancer-associated fibroblasts mediate extracellular matrix remodeling and three-dimensional invasion of scirrhous gastric carcinoma cells

Abstract Scirrhous gastric carcinoma (SGC; also called diffusely infiltrative carcinoma, Borrmann's type-IV carcinoma, or the linitis plastica-type carcinoma) have the worst prognosis among various types of gastric cancer, owing to highly progressive invasion, frequent metastasis to lymph nodes, and high incidence of peritoneal dissemination. SGC cells rapidly and diffusively infiltrate through the submucosa without forming obvious tumor mass and induce extensive stromal fibrosis, resulting in thickening and stiffening of the gastric wall. Because histophathological analyses have shown that massive proliferation of stromal fibroblasts occur within lesions of SGC, cancer-associated fibroblasts (CAFs) have been proposed to support the progression of SGC. However, biological and molecular basis of the interaction between SGC cells and CAFs remains largely unknown. In this study, we investigated the roles of CAFs in invasion and extracellular matrix (ECM) remodeling by SGC cells. When 44As3 SGC cells were co-cultured with CaF37 SGC-derived fibroblasts on 3D Matrigel, they formed large cell aggregates that invade into the Matrigel. Other SGC cell lines, HSC59 and HSC44PE, but not differentiated non-SGC gastric caner cell lines, MKN7, MKN1, and MKN74, formed invasive cell aggregate with CaF37 cells. Time-lapse imaging of SGC and CaF37 cells, and of fluorescent microsphere embedded in the Matrigel revealed that this process is associated with extensive contraction and remodeling of ECM. In order to further characterize the ECM remodeling activity of CAFs, we cultured 44As3 and CaF37 cells on fluorescent gelatin-coated coverslips. CaF37 cells caused disruption of the gelatin matrix and this activity was further promoted by co-culturing them with 44As3 cells. Immunoblotting and immnofluorescence analyses revealed that phosphorylation of myosin light chain was significantly increased in CaF37 cells co-cultured with 44As3 cells. Finally, a myosin II inhibitor blebbistatin blocked the three-dimensional invasion and ECM remodeling by 44As3 and CaF37 cells. These results indicate that SGC cells promote actomyosin-mediated contractility of stromal fibroblasts to remodel ECM during invasion. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 47. doi:1538-7445.AM2012-47

The effect of mesenchymal stem cell conditioned media on corneal stromal fibroblast wound healing activities

AimsTo investigate the effects of conditioned media from mesenchymal stem cells (MSC) on the wound healing activities of corneal stromal fibroblasts.MethodsCell cycle analysis and early stage activation of apoptosis, chemotactic...

Sensitivity of scirrhous gastric cancer to 5-fluorouracil and the role of cancer cell-stromal fibroblast interaction

Scirrhous gastric cancer is resistant to chemotherapy in comparison to other types of gastric cancers, and cancer cell-stromal fibroblast interactions play an important role in progression of scirrhous gastric cancer. We examined the effect of cancer cell-stromal fibroblast interactions on the sensitivity of an in vitro scirrhous gastric cancer model to the chemotherapeutic agent 5-FU. The model consisted of a human scirrhous gastric cancer cell line, KATO-III, and a human fibroblast cell line, KMST-6, maintained in collagen gel droplet cultures. KATO-III cell sensitivity to 5-FU was significantly decreased in co-cultures with KMST-6. KMST-6 cell sensitivity to 5-FU was also decreased in co-cultures with KATO-III. The addition of tranilast alone inhibited KMST-6, but not KATO-III, cell proliferation. In combination with 5-FU, tranilast significantly enhanced KATO-III cell sensitivity to 5-FU. These results suggest that (i), scirrhous gastric cancer cell sensitivity to 5-FU is decreased by the interactions between the cancer cells and stromal fibroblasts and (ii), tranilast enhances the sensitivity of scirrhous gastric cancer cell to 5-FU by inhibiting stromal fibroblast proliferation.

The Ets-1 transcription factor is up-regulated together with MMP 1 and MMP 9 in the stroma of pre-invasive breast cancer.

The first steps of stroma generation are of pivotal importance for carcinogenesis because at this stage are initiated both angiogenesis, the prerequisite for continuous tumour growth, and the proliferation of stromal fibroblasts. These developments contribute to the onset of tumour invasion by secreting several matrix-degrading proteases. Both angiogenesis and the production of proteases are tightly controlled at several levels; of significant importance is transcription. The Ets-1 transcription factor transactivates several genes encoding matrix-degrading proteases and is thought to be involved in both tumour vascularization and invasion. This study therefore investigated, by in situ hybridization and immunohistochemistry, the expression of Ets-1 and of two of its target genes, encoding matrix metalloproteinase (MMP) 1 and MMP 9, in order to demonstrate a topographical in vivo correlation between the expression of these three genes during breast cancer formation. All three genes were first expressed within both endothelial cells and stromal fibroblasts during the onset of stroma generation around intraductal and intralobular in situ carcinomas and they were significantly up-regulated in the stroma of invasive ductal and lobular cancers. The results of this study further support the suggested in vivo role of Ets-1 for both angiogenesis and tumour invasion, via matrix-degrading proteases which are already expressed during the early stages of breast carcinogenesis.

In vitro culture of human prostatic tissue.

A procedure is described which yields a significant percentage of long-term mixed cell cultures of human prostatic tissue. Attempts were made to suppress the proliferation of stromal fibroblasts and to characterize the cultured cells as those of prostatic origin. The problems associated with establishing epithelial cell lines are discussed.