Expression Of EGFR And ADAM9 In Pancreatic Cancer Panc-1 Cell Spheres

Objective The theory of cancer stem cells(CSCs) offer us a new thought about tumors, more and more kinds of cancer stem cells were isolated and identified form corresponding tissue. Our aim was to isolate and identify CSCs in Pancreatic cancer cell lines Panc-1 with serum-free medium(SFM)including epidermal growth factor(EGF), basic fibroblast growth factor(bFGF), insulin and B27 supplement, and to detect the expression level of EGFR and ADAM9 by Real-time PCR and Western blot.Methods Self-renewal assay, differentiation assay, detection of side population (SP) cells were performed to characterize CSCs in Panc-1 mammospheres culturing with serum-free medium. Moreover, Real-time PCR and Western blot were used to detect the expression of EGFR and ADAM9.Results Panc-1 passaged and formed mammospheres in the absence of serum. Cell spheres differentiation occurred when serum was supplemented in SFM. SP cells in Panc-1 mammospheres were(3.38±0.46)%, which showed increased expression compared with the differentiated cells (P<0.05). The expression of EGFR and ADAM9 mRNA were 2.5,3.0 times higher in mammospheres than in the differentiated cells. respectively (P<0.05). Western blot analysis suggested that EGFR and ADAM9 protein were expressed in mammospheres at higher levels than that in the differentiated cells(P<0.05).Conclusions Pancreatic cancer cell spheres may be a source for collecting CSCs by culturing with serum-free medium.ADAM9 may play an important role in the occurrence and development of Pancreatic cancer through the EGFR signaling pathway and ADAM9 is expected to become new therapeutic targets for Pancreatic cancer.