Rna Interference To Inhibit Pancreatic Cancer Cell Line Expression Of Fascin Genes

Pancreatic ductal adenocarcinoma is one of the most lethal malignancies in human. Pancreatic cancer cell has an extremely strong ability of invasion and metastasis, which is directly correlated with its poor prognosis. It is urgent for both the clinicians and the scientists who devote themselves to pancreatic cancer to find a breakthrough of curable therapy.Fascin is an evolutionary highly conserved cytoskeletal protein that is expressed in activated migrating cells and in cells with numerous membranous protrusions. Fascin is located in the core actin bundles of spikes and filopodia at the leading edge of migrating cells, and has been implicated in cell motility in several cell types playing an important role in cell migration, adhesion, and cell-cell interaction. There are several reports on fascin expression in human epithelial neoplasms. Fascin is the one of the most differentially expressed genes in pancreatic cancers compared with normal pancreas tissues using global gene expression profiling and proteomics technology. However, the role of fascin in pancreatic cancer has not clearly delineated.In our study, a human pancreatic cancer cell line, PANC-1, was transfected with the plasmid pSilencer2.1-U6 neo containing target sequence for fascin-1 gene, along with the scramble sequence as control. We found the effective target sequence for fascin-1 gene using RNA interference. Stable G418-resistant clones were obtained, and the expanded cells were then used for subsequent studies. The roles of fascin in cell behaviors such as morphology, proliferation, migration, invasion, etc. were detected, and the significance of fascin as a potential gene therapy target was evaluated.Our experiments showed that the interference of fascin expression could slow down the cell proliferation in vitro and in vivo. The proliferating cells in cell cycle might stop at G1 stage but showed scarce apoptotic cell. Fascin suppression could inhibit the formation of cell protrusion and cell microvilli as well as cause a decrease mitochondria number. Moreover, whirled laminal structure was observed in transmission EM. We revealed that fascin silencing might promote the cell adhension and reduce the anchorage-independent growth of the tumor cells in soft agar. Furthermore, interference of fascin expression could decrease the ability of cell migration and invasion. The correlative proteins detection found that fascin suppression had no effect on E-cadherin,β-catenin,β-actin or vimentin expression. The production of MMP-2 could be inhibited by fascin interfering, which showed no definite relation with MMP-9 or cathepsin D.The results of our study indicate that fascin plays an intriguing role in tumor cell morphology, proliferation, adhesion, anchorage-independent growth, migration and invasion, all of these can facilitate tumor's invasion and metastasis. Consistent with these results, fascin may be a promising target for inhibiting the invasion and metastasis of pancreatic cancer.