Nestin Delineates Pancreatic Cancer Stem Cells in Metastatic Foci of NOD/Shi-scid IL2Rγnull (NOG) Mice

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is associated with a high incidence of hepatic metastases, as well as occasional pulmonary metastases. To delineate the potential role of cancer stem cells (CSCs) in PDAC metastasis, human PDAC cells were injected into the spleen of mice. The characteristics and expression of markers associated with CSC and epithelial-mesenchymal transition (EMT) of metastatic cells that developed in the liver and lung were then compared with parental cells. The metastatic cells were polygonal, and larger than parental cells. Metastatic cells also exhibited decreased proliferation and increased adhesion to extracellular matrices, as well as enhanced migration and invasion in vitro and increased metastatic capacity in vivo. The CSC markers ALDH1A1, ABCG2, and nestin were expressed at high levels in metastatic cells and exhibited changes consistent with EMT (eg, decreased E-cadherin expression). Moreover, metastatic cells readily formed spheres in culture and exhibited an increased side population by flow analysis. Nestin and ABCG2 were also expressed at high levels in metastatic lesions from PDAC patients, and silencing nestin with shRNA in PDAC cells derived from lung metastases resulted in a marked decrease in the capacity of the cells to form spheres and to yield pulmonary or hepatic metastases. Thus, the metastatic potential of human PDAC cells correlates with CSCs and with EMT characteristics and is dependent on nestin expression.

Inhibition of nestin suppresses stem cell phenotype of glioblastomas through the alteration of post-translational modification of heat shock protein HSPA8/HSC71

“…Our previous studies have shown that nestin regulates the sphere formation ability of glioblastomas, melanomas, lung adenocarcinomas, and pancreatic cancer [23][24][25]. In this study, nestin was detected in perivascular and perinecrostic lesions, which are considered a niche for BTSCs.…”

“…We have also shown that knockdown of nestin expression suppresses migration, invasion, and sphere formation in pancreatic cancer [22,23], malignant melanomas [24] and lung cancer [25]. Similarly, Lu et al demonstrated that blocking the expression of nestin in glioblastomas by intratumoral injection of shRNA significantly reduced tumor growth and volume [19].…”

Electron microscopic analysis of different cell types in human pancreatic cancer spheres

“…In a previous study, we reported that nestin, a pancreatic CSC marker, is more highly expressed in the spheres of three PDAC cell lines than in nonsphere cells (13). Moreover, pancreatic cancer cells derived from metastatic foci of immunodeficient mice form a greater number of spheres in low-attachment plates than do their primary tumor counterparts (14). These findings suggest that CSCs are enriched in the spheres and play an important role in the aggressiveness of PDAC.…”

Systemic Administration of Small Interfering RNA Targeting Human Nestin Inhibits Pancreatic Cancer Cell Proliferation and Metastasis

“…17 After 5 days of culture, the number of spheres in each well was counted. Total RNA was extracted from spherical cells and nonspherical cells and was used for PCR analysis.…”

“…16,17 Immunoblotting was performed using an anti-nestin antibody (diluted 1:2,000; BD Bioscience, Franklin Lakes, NJ).…”