TY - JOUR
T1 - Prometastatic mechanisms of CAF-mediated EMT regulation in pancreatic cancer cells
AU - Shan, Tao
AU - Chen, Shuo
AU - Chen, Xi
AU - Lin, Wan Run
AU - Li, Wei
AU - Ma, Jiancang
AU - Wu, Tao
AU - Ji, Hong
AU - Li, Yiming
AU - Cui, Xijuan
AU - Kang, Yaan
PY - 2017/1
Y1 - 2017/1
N2 - Tumor metastasis are accompanied by the EMT (epithelial-mesenchymal transition)-MET (mesenchymalepithelial transition) two-step process. In this study, we investigated the importance of cancer associated fbroblasts (CAF) in the process. First, the primary cultures of isolated pancreatic CAF, fbroblasts of normal pancreatic tissues (NF), and normal hepatic stellate cells (HSF) were identified and verified via the expression of a-SMA and vimentin. Using an indirect three-dimensional co-culture model, the morphological changes were observed by light microscopy and laser scanning confocal microscopy. The invasive and migration capacity of pancreatic cancer cells was determined by Transwell chamber migration assay or scratch assay. The mRNA and protein expression levels of E-cadherin, vimentin, and Gli1 were determined by RT-PCR and western blotting. Primary cultures of isolated CAF, NF, HSF showed satisfactory growth with active proliferation. Indirect co-culture with CAF, BxPc-3 and Panc-1 cells showed significant partial EMT, reduced E-cadherin expression, and enhanced vimentin expression as compared with the single culture and NF/HSF co-culture groups, with corresponding increases in migratory and invasive capacities. PCR and western blotting results showed that mRNA and protein expression levels of Gli1 in CAF and Snail in cancer cells were increased. This process could be reversed by inhibition of hedgehog (HH) signaling in CAF. In the tumor microenvironment, activation of CAF is the key event in mediating partial EMT, and its mechanism may be associated with paracrine action after activation of HH signaling in CAF.
AB - Tumor metastasis are accompanied by the EMT (epithelial-mesenchymal transition)-MET (mesenchymalepithelial transition) two-step process. In this study, we investigated the importance of cancer associated fbroblasts (CAF) in the process. First, the primary cultures of isolated pancreatic CAF, fbroblasts of normal pancreatic tissues (NF), and normal hepatic stellate cells (HSF) were identified and verified via the expression of a-SMA and vimentin. Using an indirect three-dimensional co-culture model, the morphological changes were observed by light microscopy and laser scanning confocal microscopy. The invasive and migration capacity of pancreatic cancer cells was determined by Transwell chamber migration assay or scratch assay. The mRNA and protein expression levels of E-cadherin, vimentin, and Gli1 were determined by RT-PCR and western blotting. Primary cultures of isolated CAF, NF, HSF showed satisfactory growth with active proliferation. Indirect co-culture with CAF, BxPc-3 and Panc-1 cells showed significant partial EMT, reduced E-cadherin expression, and enhanced vimentin expression as compared with the single culture and NF/HSF co-culture groups, with corresponding increases in migratory and invasive capacities. PCR and western blotting results showed that mRNA and protein expression levels of Gli1 in CAF and Snail in cancer cells were increased. This process could be reversed by inhibition of hedgehog (HH) signaling in CAF. In the tumor microenvironment, activation of CAF is the key event in mediating partial EMT, and its mechanism may be associated with paracrine action after activation of HH signaling in CAF.
KW - Cancer associated fibroblasts
KW - Epithelial-mesenchymal transition
KW - Hedgehog signaling
KW - Pancreatic cancer
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U2 - 10.3892/ijo.2016.3779
DO - 10.3892/ijo.2016.3779
M3 - Article
C2 - 27878234
AN - SCOPUS:85007603317
SN - 1019-6439
VL - 50
SP - 121
EP - 128
JO - International journal of oncology
JF - International journal of oncology
IS - 1
ER -