摘要目的 观察肝素酶与非阳离子依赖型甘露醇-6-磷酸受体(CD222)在膀胱癌5637细胞中的结合及两者结合后对细胞侵袭性的影响.方法 应用免疫荧光双染方法研究肝素酶与CD222在膀胱癌5637细胞中定位及结合.设计、合成小干扰RNA(siRNA)-CD222,转染膀胱癌5637细胞.逆转录-聚合酶链反应(RT-PCR)、Western blot法检测转染效率.加入人重组肝素酶后采用Western blot法检测转染前后CD222及磷酸蛋白激酶B(p-Akt)的表达.Transwell侵袭实验检测以上细胞的侵袭性.结果 肝素酶与CD222均表达于细胞质.加入人重组肝素酶后5637细胞CD222、p-Akt的表达量及侵袭性显著增强为(79.71 ±3.25)个,而上述效应在转染siRNA-CD222后明显降低为(49.61 ±2.88)个,差异有统计学意义(P＜0.05).结论 5637细胞中肝素酶可与CD222结合,两者共同定位于细胞质.肝素酶通过与其受体CD222细胞外结合可促进Akt磷酸化,进一步促进膀胱癌5637细胞的侵袭.

abstractsObjective To study the relationship between heparanase and cation-independ mannose 6-phosphate receptor (CD222) expression in bladder cancer 5637 cell line.And find out the effect of cell invasiveness by their combination.Methods Immunofluorescence assay were used to detect the position fixing and interaction of heparanase and CD222 in bladder cancer 5637 cell line.The CD222 mRNA-targeted small interfering RNA (siRNA) were designed and constructed.5637 cells were cultured and transfected with siRNA-CD222.The expression of CD222 mRNA and protein were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting.After the recombinant human heparanase were given.Western blotting were performed to detect the expression of CD222 and p-Akt in 5637 cell line before and after tansfection.The invasiveness of above cells were determined by matrigel invasion assays.Results Heparanase and CD222 staining on the cell cytoplasm.Heparanase binds CD222 and cells treated with recombinant human heparanase enhance the binding.Inhibition of CD222 expression in 5637 by siRNA could decrease invasion capacity and the level of Akt phosphorylation which was induced by recombinant human heparanase.Conclusion Heparanase binding to CD222 were localized to the cell cytoplasm and heparanase facilitates bladder cancer cell invasiveness by binding to CD222 and increasing Akt phosphorylation.