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目的：通过构建过表达质粒并筛选稳定细胞株，研究SPATA5L1基因对KB细胞增殖和迁移的影响。方法 PCR扩增SPATA5L1基因全长，用载体pEX-1构建重组质粒，测序成功后转染人口腔癌KB细胞，并设空载体作对照，经过G418压力筛选后获得单克隆细胞株，Real-time PCR、Western blot验证SPATA5L1基因的表达，光镜观察SPATA5L1对KB细胞形态的影响，CCK-8法检测细胞增殖情况，划痕实验观察细胞迁移能力。结果测序结果显示质粒构建成功，经过G418压力筛选所得到的稳定表达细胞株，在转录水平和蛋白水平均检测到目的基因的高表达，且细胞由正常状态下的多边形变为梭形，细胞增殖能力和体外迁移能力明显提高（P＜0.05）。结论成功构建pEX-1-SPATA5L1表达载体并获得稳定细胞株，过表达SPATA5L1蛋白能使KB细胞形态变长，增殖和迁移加快。“,”Objective To construct the recombinant eukaryotic vector expressing SPATA5L1 cDNA,and explore the effect of SPATA5L1 over-expression on the proliferation and migration of human oral cancer cell line KB.Methods The fragment of SPATA5L1 gene was ampliifed by polymerase chain reaction(PCR).An eukaryotic vector expressing SPATA5L1(pEX-1-SPATA5L1) was constructed and transfected into KB cells which were then selected in culture medium containing G418 to acquire the monoclonal cell lines.The transcription and expression of SPATA5L1 gene were detected by Western blotting.The effect on the proliferation and migration of KB cells were evaluated by CCK-8 assay and wound healing assay respectively.Results The plasmid vector with target SPATA5L1 gene was designed correctly.The monoclonal cell line which showed spindle shape was obtained after screened with G418.The protein level of SPATA5L1 in the pEX-1-SPATA5L1 transfected cells was signiifcantly higher than controls.CCK-8 assay showed that the proliferation of pEX-1-SPATA5L1-KB cells was much faster than the controls.Wound healing assay revealed that up-regulating SPATA5L1 in KB cells could promote the wound healing.Conclusion Up-regulating SPATA5L1 gene could stimulate the proliferation of KB cells and promote the migration of KB cells.