Biochemical and Biophysical Research Communications
Role of fascin in the proliferation and invasiveness of esophageal carcinoma cells☆
Section snippets
Materials and methods
Materials and cell culture. Human ESCC cell line EC109 and uterine cervix cancer cell line HeLa were purchased from the Chinese Academy of Science. They were maintained in 199 medium (Invitrogen) containing 10% fetal calf serum (FCS) in an atmosphere of 5% CO2 at 37 °C. Monoclonal antibody anti-fascin, anti-PCNA, anti-c-erbB-2 (Dako Corporation), anti-β-catenin (Santa Cruz), anti-β-actin (Sigma), and anti-topoisomerase II (Roche) were used for immunofluorescence and Western blotting. Anti-mouse
Fascin expression in EC109 cells
By using Western blotting, we first examined the level of fascin expression in EC109 cells and HeLa cells. Fascin expression in EC109 cells is almost as high as that in HeLa cells (a putative fascin overexpression tumor cell line), indicating that fascin was also upregulated in EC109 cells (Fig. 1A). Immunofluorescence showed that fascin was localized in the cytoplasm, especially in the membrane protrusions of EC109 cells (Fig. 1B).
Silencing of the fascin gene
To study the function of fascin gene in EC109 cells, pSUPER
Discussion
Fascin is an actin-bundling protein originally found in the extracts of unfertilized sea urchin eggs [2]. Fascins play roles in the organization of two major forms of actin-based structures: dynamic cortical cell protrusions and cytoplasmic microfilament bundles. High-level expression of fascin is observed in many tumor cell lines such as breast carcinoma and esophageal carcinoma, but the roles and mechanism still remained unclear. For a better understanding of the function of fascin, in the
Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (No. 39900069, No. 30170428, and No. 30370641); Guangdong Scientific Fund Key Items (No. 37788); Natural Science Foundation of Guangdong Province (No. 010431). We thank Dr. S.J. Fang for the kind help in siRNA design and Professor X.J. Yu for his kind assistance in photomicrography.
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2016, Experimental Cell ResearchCitation Excerpt :Fascin is increasingly recognized to play some, even not fully understood, critical functions in cytoskeleton remodeling. Namely, decreased level of fascin correlated with decreased formation of dynamic cell protrusions, decreased proliferation, and reduced invasiveness in cancer cells [35,36]. As expected, we observed a marked decrease in fascin immune staining in INS-treated cells along with the aforementioned sharp decrease in lamellipodia and filopodia (Fig. 8a).
TGF-β1-induced Fascin1 promotes cell invasion and metastasis of human 786-0 renal carcinoma cells
2016, Acta HistochemicaCitation Excerpt :According to the follow-up experiment, the cells were divided into six groups including blank control group (also NT group, non-transfected 786-0 cells), negative control group (also si-NC group, 786-0 cells transfected with pGenesil-1-con plasmid), positive group (also si-Fascin1 group, 786-0 cells transfected with pGen-1-FSCN1 plasmid), and three corresponding groups: NT treated with TGF-β1, si-NC treated with TGF-β1 and si-Fascin1 treated with TGF-β1 (Chen, 2014a,b). Based on the method used in the study of Xie et al. (2005), the interference sequence of Fascin1 genes was effectively screened. The most complete coding sequence of Fascin1 gene was found in GenBank by means of bioinformatics methods.
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Abbreviations: ESCC, esophageal squamous cell carcinoma; RNAi, RNA interference; siRNA, short interfering RNA; FCS, fetal calf serum; TBS, Tris-buffered saline; SEM, scanning electron microscopy; PCNA, proliferating cell nuclear antigen; MMP-2 and MMP-9, matrix metalloproteinases-2 and -9.
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These authors contributed equally to this work.