Establishment and characterization of a carcinoma-associated fibroblast cell line derived from a human salivary gland adenoid cystic carcinoma

Cancer-associated fibroblasts (CAFs) are thought to play an essential role in cancer initiation and development. However, little research has been done to evaluate the role of CAFs in epithelial ovarian cancer (EOC) development. To address this issue, ninety-one specimens were immunostained with α-smooth muscle actin (α-SMA) and fibroblast activation protein (FAP) antibodies to quantify CAFs, and antibodies D2-40 and CD34 to evaluate the lymphatic vessel density (LVD) and microvessel density (MVD) of the lesions. We found there were no α-SMA or FAP positive fibroblasts in normal ovary tissues. More CAFs were found in EOC than in borderline tumors and benign tumors (P<0.01). Abundant CAFs in EOC were associated with advanced-stage disease (P=0.002), the occurrence of lymph node metastases (P=0.02) and omentum metastases (P<0.0001), and increased LVD (P=0.002) and MVD (P=0.0004). CAFs isolated from EOC tissues induced more cancer cells to invade (P=0.003) and migrate (P=0.005) compared with normal fibroblasts (NFs) isolated from normal ovary tissues in vitro. Our data indicate that CAFs play a vital role in ovarian cancer progression and metastasis. Targeting CAFs as a therapeutic strategy against ovarian cancer is an intriguing concept that needs further study. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Cancer-associated fibroblasts (CAFs) are reported to support tumorigenesis by stimulating angiogenesis, cancer cell proliferation, and invasion in most solid tumors. However, the roles of CAFs in the liver cancer microenvironment have not been thoroughly studied. In our previous study, we successfully isolated CAFs from hepatocellular carcinoma (HCC) (H-CAFs) and proved that H-CAFs suppressed the activation of NK cells and thereby created favorable conditions for HCC progression. In our present study, we found that the proliferation of MHCC97L and Hep3B cells was significantly promoted by treatment with conditioned medium from H-CAFs. Pathological analysis also revealed that H-CAFs increased the proportion of Ki-67 (+) malignant cells and prevented them from undergoing necrosis. Moreover, the concentration of hepatocyte growth factor (HGF) cytokine in the conditioned medium of H-CAFs was higher than conditioned medium from normal skin fibroblasts (NSFs). Anti-HGF significantly reduced the proliferation-promoting capability of H-CAFs. In addition, we found that the abundance of H-CAFs correlated positively with tumor size. These results indicate that H-CAFs are an important factor for promoting the growth of HCC in vitro and in vivo, and that HGF plays a key role in HCC proliferation induced by H-CAFs.

Carcinoma-associated fibroblasts (CAFs) are a key determinant in malignant progression of cancer and represent an important target for cancer therapies. In this work, we present a microfluidic-based 3D co-culture device to reconstruct an in vitro tumor microenvironment and firstly investigate the effect of CAFs on cancer cell invasion in 3D matrix. This device is composed of six co-culture units, which enable parallel co-culture assays to be run in the presence of 3D extracellular matrix. Salivary gland adenoid cystic carcinoma (ACC) cells and CAFs embedded in matrix were co-cultured without direct contact on the device. Communication between ACC cells and CAFs could be established via medium diffused in matrix. It was observed that CAFs promoted ACC cell invasion in 3D matrix in a spheroid fashion, indicating that CAFs play a critical role in cancer invasion. We further demonstrated the effect of MMP inhibitor as an agent against CAF-promoted cancer invasion. This co-culture device reproducibly reflected the in vivo growth and invasion pattern of ACC and recreated the stroma-regulated ACC invasion. Thus, it provides a suitable platform for elucidating the mechanism of CAF-regulated cancer invasion and discovering anti-invasion drugs in a well defined 3D environment.