【摘要】 目的研究L-选择素(L-selectin)与其配体甘露糖受体(mannose receptor,MR)间相互作用与小鼠肝癌细胞Hepa1-6淋巴道转移潜能相关性。方法构建含有L-selectin的重组载体pcDNA3.1,转染小鼠肝癌细胞Hepa1-6,筛选稳定表达L-selectin的细胞株。采用免疫组化、RT-PCR、流式细胞、体外黏附试验技术研究小鼠肝癌细胞Hepa1-6中L-selectin的表达及其与肿瘤细胞淋巴道转移潜能的相关性。结果 1重组质粒为目的基因L-selectin。2 Hepa1-6细胞表面有L-selectin表达,阳性率20.2%。3流式细胞仪检测结果表明Hepa1-6细胞表达L-selectin为19.9%。4体外黏附试验结果表明,不具有转移特性的Hepa1-6表达L-selectin后可发生淋巴道转移,L-selectin能与MR结合(P<0.001),介导Hepa1-6的淋巴道转移。结论 Hepa1-6表达目的基因L-selectin后具有淋巴道转移潜能,并能与MR在体外结合,介导肿瘤细胞的淋巴道转移。更多还原

【Abstract】 Objective To study the correlation between L-selectin and its ligand mannose receptor with mice hepatoma cells Hepa1-6 lymphatic metastatic potential.Methods The recombinant plasmids pcDNA3.1 containing L-selectin was constructed,and transfected mouse liver tumor Hepa1-6 cells.The cell strains with a stable expression of L-selectin were screened.Immunochemistry,flow cytometric analysis,RT-PCR and other procedures,in addition to in vitro adhesion assays were performed,to verify the expression of L-selectin on the surface of Hepa1-6 cells,and the interactions between L-selectin and tumor cells lymphatic metastasis potential was studied.Results The target gene of recombinant plasmid was L-selectin.L-selectin was expressed on Hepa1-6 cell’ s surface with the positive rate of 20.2%.By flow cytometric analysis,the positive expression rate of L-selectin in Hepa1-6 cells was 19.9%.In vitro adhesion assays indicated that this protein was able to bind specifically to MR(P<0.001),and further confirmed that Hepa1-6 cells without metastatic properties were capable of lymphogeneous metastasis following the expression of L-selectin.Conclusion The recombinant expression vector for L-selectin is successfully expressed in Hepa1-6 cells.L-selectin expressed on the surface of Hepa1-6 cells was fully functional,capable of binding to MR in vitro and mediating the tumor cell lymphatic metastasis.更多还原