| Objective: Our study aims at the expression of Siglec-15 in giant cell tumor of bone(GCTB),and investigating the correlation between Siglec-15 in GCTB tissues and clinical data.Additionally,we examed the expression of Siglec-15 in GCTB stromal cells,Hs 737.T cells.si RNA was used to observe the influence of Siglec-15 on the biological characteristics of Hs 737.T cells such as proliferation,migration and invasion in vitro.Methods: Fifty-six specimens of GCTB were gathered from the Department of Pathology,the Third Hospital of Hebei Medical University from January 1,2012 to December 31,2017.Immunohistochemical assay was used to exam Siglec-15 protein in GCTB tissues.The association between Siglec-15 expression and clinical information such as gender,age,location,size,Companacci’s staging,and tumor recurrence was analyzed.The expression of Siglec-15 in GCTB stromal cells was determined by RT-PCR and immunofluorescence technique.The gene expression of Siglec-15 in Hs737.T cells was silenced by si RNA.The decreased levels on m RNA and protein of Siglec-15 by si RNA were certificated by RT-PCR and Western blot.The functions of growth,clone formation,migration and invasion of Hs 737.T cells were investigated by the experiments of proliferation,clone formation,wound scratch and Transwell.Results: Immunohistochemical assay showed that Siglec-15 mainly located in the cytoplasm and membrane of cells in GCTB tissues.In 56 specimens,51 were positive for Siglec-15(91.07%).Siglec-15 positive expression in Companacci’s staging and tumor recurrence had significant difference(P<0.05).There was no significant difference of Siglec-15 expression in gender,age,location and size.Immunofluorescence staining demonstrated that Siglec-15 was largely expressed in the cytoplasm of Hs737.T cells.The RT-PCR showed that Siglec-15 m RNA was highly expressed in Hs 737.T cells(1.636±0.012).The RT-PCR showed that there was significantly reduced Siglec-15 expression transfected with si RNA for 36 h in comparison to negative control group.The Western Blot showed there was obviously reduced Siglec-15 expression transfected with si RNA at 48 h.The functions of proliferation,migration and invasion of Hs 737.T cells all reduced with si RNA transfection(P<0.05).Conclusion: Siglec-15 is highly expressed in GCTB tissues,and has significant difference in Companacci’s staging and tumor recurrence.The functions of proliferation,migration and invasion of Hs 737.T cells significantly decrease with Siglec-15 reduction.These suggest Siglec-15 plays an important role in the occurrence and development of GCTB,and may be a new indicators for the diagnosis,treatment and prognosis of GCTB. |
