转染野生型DNA聚合酶β基因对交链孢霉酚毒性作用的影响  

A Study of DNA Polymerase β Gene Mutations Induced by AOH

作　　者：徐华明[1] 张文靖[1] 

机构地区：[1]河南中医学院,河南郑州450008

出　　处：《肿瘤基础与临床》2007年第4期297-298,共2页journal of basic and clinical oncology

摘　　要：目的观察转染野生型DNA聚合酶β(DNApolβ)基因对交链孢霉酚(AOH)诱导NIH3T3细胞DNApolβ表达量改变的影响。方法用pEGFP-C3、pEGFP-C3-polβ质粒转染NIH3T3细胞;将转染pEGFP-C3和pEGFP-C3-polβ的NIH3T3细胞分别用1.5μmol/L、2.0μmol/L和2.5μmol/L3种浓度的AOH诱导48 h后,RT-PCR同时扩增DNA polβ基因和β-actin基因,进行半定量分析。结果在AOH作用下,横向比较发现,3种细胞鼠polβ表达量都有升高的现象,且AOH浓度越高,表达量升高的越明显;但经纵向分析发现,在同样浓度的AOH诱导下,转染野生型的polβ的NIH3T3细胞的鼠polβ升高的程度较未转染的NIH3T3细胞和转染空载体的NIH3T3细胞鼠的表达量低(P<0.05)。结论高表达野生型DNA polβ具有一定程度的抵抗霉菌毒素这种诱变剂致使NIH3T3细胞DNA polβ表达量升高的作用。Objective Tq observe the effect of AOH on the expression levels of polymerase β, after transfecting the plasmid of wild DNA polβ. Methods The vectors of pEGFP-C3 and pEGFP-C3-polβ were transfected into NIH3T3 cell lines with lipofectamine method, and screened with C,418, two stably NIH3T3 ,cell lines were obtained carrying pEGFP-C3-polβ and pEGFP-C3, respectively. The two groups of NIH3T3 cells （carrying pEGFP-C3 and carrying pEGFP-C3-polβ） were induced for 48 hours by the following concentrations of AOH： 1.5 μmol/L, 2.0μmol/L, 2.5 μmol/L. Total RNA were extracted and reverse transcripted to cDNA. Semi-quantitative PCR were performed to detect the expression levels of DNA polβ, where β-actin was performed as inner control, in NIH3T3 cells carrying different vectors. Results The expression levels of DNA poll3 increased as the AOH concentration increased in all the cells. But in NIH3T3 cells carrying pEGFP-C3 and untransfected NIH3T3 cells, the expression level of DNA poll3 were obviously higher than those of NIH3T3 cells carrying pEGFP-C3- polβ on the same concentration of AOH. Conclusions To some extent, overexpressive wild DNA polymerase β can inhibit the effect of AOH on the expression levels of polymerase β.

关 键 词：DNA聚合酶Β 霉菌毒素 半定量PCR 

分 类 号：R730.231[医药卫生—肿瘤]