The Research Of Capturing CXCR4 In Head And Neck Squamous Carcinoma Cells To Inhibit Lympatic Metastasis

ObjectivesTo study the effects of CXCL12-KDEL fusion gene on the growth and lymph metastasis of the implanted carcinoma in nude mice after transfected into human oral squmaous carcinoma cell(OSCC) line Tb3.1.To explore the capturing effect of CXCL12-KDEL fusion gene to CXCR4.To find out a effective therapy target restraining the proliferation and metastasis and make a initial approch of target therapy of OSCC.MethodsCXCL12-KDEL-pIRES2-EGFP fusion gene and eukaryotic vector pIRES2-EGFP were electric transfected into Tb3.1 cell line.After G418 selection and single cell clone proliferation,we obteined stable transfected cell line CXCL12-KDEL-pIRES2-EGFP-Tb3.1 and pIRES2-EGFP-Tb3.1. Establish the tongue cancer cell nude mice model, the mice mucosa were injected with 5×105 cells. observe the the growth of tumor and draw the curve of tumor volume. Then take out the tumor and lymph, nodes,HE and IHC staining analysis the changing of pathology and expression of proteins related with metastasis and proliferation.Results1. CXCL12-KDEL-pIRES2-EGFP fusion gene and eukaryotic vector pIRES2-EGFP were electric transfected into Tb3.1 cell line.After G418 selection and single cell clone proliferation,we obteined stable transfected cell line CXCL12-KDEL-pIRES2-EGFP-Tb3.1 and pIRES2-EGFP-Tb3.1.2. Establish the tongue cancer cell nude mice modelThe tumor were formed after injected with 5×105 cells with 7days.4 weeks later, take out the tumor, measure the volume,there was a significant difference between the three groups(P<0.05). HE stain show the CXCL12-KDEL-pIRES2-EGFP group has no lymph metastasis,other two groups have 8 and 9 (P<0.05).3.The histopathologic change and expression of special protein related with metastasis and proliferation in Tb 3.1 nude mice model.The expression rates of Cox-2 in the CXCL12-KDEL-pIRES2-EGFP group pIRES2-EGFP group and the non-transfected group were 70%(21/30),90%(36/40),87.88%(29/33),there were stastical differences among the three groups(P<0.05).The expression rates of Bcl-2 in the CXCL12-KDEL-pIRES2-EGFP group pIRES2-EGFP group and the non-transfected group were 50%(8/16),84%(21/25),85.71%(18/21),there were stastical differences among the three groups (P<0.05).The expression rates of VEGF-C in the CXCL12-KDEL-pIRES2-EGFP group, pIRES2-EGFP group and the non-transfected group were 54.55%(12/22),85.20% (23/27),83.16%(16/19),there were stastical differences among the three groups (P<0.05).ConclusionCXCL12-KDEL-pIRES2-EGFP fusion gene can capture CXCR4,block the CXCL12-CXCR4 axis could inhibit the cell prolification and lymph node metastasis. CXCR4 may be an effective target of the therapy of OSCC.