不同类型的DNA聚合酶β基因对CHO细胞的影响

Effect of wild-type and mutant DNA polymerase β expression on CHO cells growth

目的:比较野生型和突变型(58 bp缺失)DNA聚合酶β(polβ)基因对真核细胞生长特性的影响.方法:将表达人野生型和缺失型DNA polβ的中国仓鼠卵巢细胞株(CHO),用RT-PCR方法鉴定野生型和缺失型DNA polβmRNA水平的表达,MTT法测生长曲线、流式细胞术测细胞周期.结果:转染野生型和突变型DNA polβ的CHO细胞株中,都有外源基因mRNA水平的表达;转染突变型(58 bp缺失)polβ基因的细胞增殖速度和S期比例增高均高于未转染组细胞,差异有统计学意义(P<0.05).而转染野生型polβ基因对细胞增殖速度和细胞周期无明显影响(P>0.05).结论:高表达外源性野生型和突变型DNA polβ,对CHO细胞生长具有不同的影响.

AIM： To study the effects of the DNA polymerase 13 （DNA poll3） of wild type and the mutant（58 bp deletion） on Chinese hamster ovary cells （CHO）. METHODS： The expression of mRNA of wild type and the mutant in transfectant cells was determined by RT-PCR and the growth curve of transfected CHO cells was drawn with the MTT assay. The changes in cell cycle were analyzed by flow cytometry. RESULTS： Cell lines stably expressing wild-type and the mutational polβ were established. The CHO cells transfacted by the mutant gene were higher than untransfacted CHO cells in the growth rate of CHO cells and the percentage of S phase cells in vitro （ P 〈 0.05 ）. The effects of the wild-type enzyme on growth rate and cell cycle were not observed （ P 〉 0.05 ）. CONCLUSION： The wild type and the mutant have different effect on CHO cells.