The Preliminary Study On The Relationship Between CD133~+ Cancer Stem-Like Cells And Lymphangiogenesis In OSCC

objective:Oral squamous cell carcinoma(OSCC)is the most common histological type of oral malignancy,and due to its prone to lymph node metastasis,patients with OSCC have poor therapeutic and prognostic effects,thus receiving high attention from researchers.One of the reasons for the cancer refractory is cancer stem cells.This study will analyze the cancer stem-like cells’characteristics through the detection of cancer stem cell markers CD133 in OSCC,and preliminary study the relationship between the CD133~+cancer stem-like cells of OSCC and lymphangiogenesis as well as the possible mechanism,Proposed to provide new direction for improving the treatment and prognosis of OSCC.Methods:1.163 cases of paraffin tissue specimens of OSCC and37 cases of normal oral mucosal tissue were collected.Immunohistoch-emistry was used to detect the expression of CD133 protein in OSCC and normal oral mucosa tissues,and to analyze the relationship between its expression and the clinicopathological parameters of patients with OSCC,the prognostic value of the CD133’s expression was also analyzed.2.The expression of VEGF-C protein in OSCC and normal oral mucosa tissues was detected by immunohistochemistry,and analyzed the the correlation with the clinicopathological parameters of patients with OSCC as well as the expression of CD133 protein.3.The expression of podoplanin protein in OSCC and normal oral mucosal tissue was detected by immunohistochemistry,and analyzed the relationship between lymphatic vessel density and the expression of CD133 protein(lymphatic vessel density was calculated by the expression of podoplanin protein).4.The expression of the markers for lymphatic vessel endothelial cell(LYVE-1,Podoplanin and VEGFR-3)in CD133~+SCC9 human tongue squamous cell carcinoma(after the induced differentiation of recombinant human VEGF-C)were detected by western-blot and qPCR in protein and gene level,to explore the the relationship between the CD133~+cancer stem-like cells of OSCC and lymphangiogenesis as well as the possible mechanism.Results:1.The expression of CD133 protein in OSCC was higher than that in normal oral mucosal(P<0.05),and its expression was related to the clinical stage,differentiation degree,lymph node metastasis and prognosis of OSCC(P<0.05),and may be an independent factor for the prognostic of OSCC patients.2.VEGF-C expression rate and lymphatic vessel density in OSCC were higher than those in normal oral mucosa tissues(P<0.05),which were related to clinical stages and lymph node metastasis of OSCC patients(P<0.05).3.Expression of VEGF-C and lymphatic vessel density in the group with positive expression of CD133 were higher than those with negative expression of CD133(P<0.05).4.The expression intensity of CD133 protein was positively correlated with VEGF-C(P<0.05).5.The expression of LYVE-1,Podoplanin and VEGFR-3(markers of lymphatic endothelial cells)in CD133~+SCC9 human tongue squamous cell carcinoma was increased under the induced differentiation culture with recombinant human VEGF-C(P<0.05).Conclusion:1.CD133 protein is highly expressed in OSCC than normal oral mucosa,and its expression is closely related to lymph node metastasis and prognosis of OSCC patients,and may be an independent factor affecting the prognosis of OSCC patients.2.The VEGF-C protein’s expression and LVD in OSCC are both higher than normal oral mucosa,and both related to the expression of CD133,suggesting that CD133~+cancer stem-like cells may be related to lymphangiogenesis in OSCC,and VEGF-C protein may be involved in the regulation of this process.3.CD133~+cancer stem-like cells may gradually differentiate into lymphatic endothelial cells after induced differentiation culture of recombinant human VEGF-C in OSCC.