B7-1基因转染对骨肉瘤细胞生物学行为的影响

目的:研究B7-1真核表达载体pcDNA3-B7-1转染对骨肉瘤细胞株LM8的生物学行为的影响。方法:用脂质体(lipofectamine)介导B7-1基因和空载体pcDNA3转染LM8细胞,G418筛选得到抗性克隆,分别命名为LM8/B7-1和LM8/pcDNA3,用聚合酶链反应(RT-PCR)和流式细胞仪(FCM)鉴定转染基因的表达,采用四甲基偶氮唑蓝微量酶反应比色法(MTT)和FCM检测细胞体外增殖活性,并观察转染细胞在C3h小鼠体内致瘤能力。结果:RT-PCR和FCM显示B7-1基因在骨肉瘤细胞株LM8中呈高表达,LM8/B7-1体外增殖活性与LM8/pcDNA3和LM8细胞差异并无显著性(P>0.05),但LM8/B7-1的致瘤能力显著下降(P<0.01)。结论:B7-1基因对骨肉瘤细胞的体内增殖能力产生明显抑制作用,可以作为骨肉瘤基因治疗的理想目的基因之一。

Biologic behavior effects of introduction of B7-1 cDNA into osteosarcoma cell line LM8

Purpose:To study the effects on biologic in cells obtained from osteosarcoma cell line LM8 into which the B7-1 cDNA was introduced. Methods:B7-1 gene was transfected into osteosarcoma LM8 cells with lipofectamine mediation.The positive clones were selected by G418 .The effects of B7-1 gene on the osteosarcoma LM8 cells were examined by using RT-PCR,flow cytometry analysis(FCMA),MTT assay and mice inoculation test.Results:The RT-PCR and FCMA showed that B7-1 was highly expressed.The oncogenicity of LM8/B7-1 cells was decreased greatly compared with B7-1-LM8(LM8) cells(P<0.01).But their ability of proliferation in vitro had no significant difference(P>0.05). Conclusions:The B7-1 gene can effectively promote the activity of CTL against osteosarcoma cells,and may be considered as one of the target genes for the gene therapy of osteosarcoma.