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Expression and secretion of nerve growth factor in esophageal squamous cell cancer cell line Eca109
Pages: 348-353
Year: Issue:  3
Journal: Acta Anatomica Sinica

Keyword:  Esophageal squamous cell carcinomaNerve growth factorEca109 cellImmunohistochemistryRT-PCRWestern blottingELISA;
Abstract: Objective To investigate the relationship between the nerve growth factor( NGF) and the cell differentiation of esophageal squamous cell cancer. Methods Round and spindle single-cell clones were sorted by limited dilution,and spheres were obtained by cultured in serum-free medium. Adherent Eca109 cells were cultured in serumcontaining and serum-free culture medium 48 hours,respectively. The expression of NGF mRNA and protein levels in esophageal squamous cell carcinoma( ESCC) were assessed by reverse transcriptase expression of the polymerase chain reaction( RT-PCR) technology and Western blotting analysis,respectively. Immunohistochemical staining determined the nerve growth factor expression in esophageal squamous cell carcinoma specimen. The secretion of NGF in Eca109 was determined by enzyme-linked immunosorbent assay( ELISA). Results The expression of NGF mRNA and protein levels was detected in ESCC,and the spheres cells expressed the highest amount of NGF in mRNA and protein expression level.NGF was expressed in the cytoplasm of ESCC cells in esophageal squamous cell carcinoma specimen. ESCC cells could secreted NGF in serum-free culture conditions,and the NGF secreted was significantly higher than that in serum-containing medium. Conclusion ESCC cells of Eca109 express and secrete NGF,and esophageal squamous cell carcinoma specimen could express nerve growth factor. NGF may play an important role in maintaining ESCC stem cell characteristics.
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