| Objective To investigate the effects of the nerve growth factor to the expression of p75 NTR in the cell nucleus of esophageal squamous cell cancer.Methods Round and spindle single-cell clones were sorted by limited dilution, and spheres were obtained by cultured Eca109 in serum-free medium, respectively. Adherent Eca109 cells were cultured in serum-containing and serum-free culture medium for 48 h, respectively. Immunohistochemical staining determined the nerve growth factor expression in esophageal squamous cell carcinoma specimen.The expression of NGF m RNA and protein levels in the cells treated above were assessed by reverse transcriptase expression of the polymerase chain reaction(RT-PCR) technology and Western blot analysis,respectively. The secretion of NGF in the cells treated above, was determined by enzyme-linked immunosorbent assay(ELISA).Results NGF was also expressed in the cytoplasm of OESCC cells in esophageal squamous cell carcinoma specimen.The expression of NGF m RNA and protein levels could be detected in all ESCC treated above,and the spheres cells expressed the highest amount of NGF in m RNA and protein level. NGF could be secreted by ESCC cells in serum-free culture medium, and the capacity of NGF secretion was significantly higher than that in serum-containing medium. After NGF treatment, The proportion of p75 NTR expression in nucleus in Eca109 and sphere cells increased. And the expression of Itgα6, Itgβ4 and Itgβ1 expressing lever was higher in sphere cells than in Eca109 cells,which was consistent with the increase of proportion of the expression of p75 NTRin nucleus.Conclusion ESCC cell could produce and secret NGF, the spheres cells expressed the highest amount of NGF,and NGF facilitated the p75NTR-ICD translocation to the nucleus. And Itgα6, Itgβ4 and Itgβ1 could be useful to identify the ESCC stem cell with p75 NTR. NGF may play an important role in the maintenance of ESCC stem cell characteristics. |
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