DNA polβ启动子和CMV启动子调控的p53基因在涎腺腺样囊性癌细胞中表达的比较  

Expression of p53 Gene with DNA polβ and CMV Promoter in Salivary Adenoid Cystic Carcinoma Cells

作　　者：阎炳智[1] 王洁[1] 张波[2] 董福生[3] 侯琳[2] 王旭[1] 

机构地区：[1]河北医科大学口腔医学院病理教研室,河北石家庄050017 [2]北京大学医学院病理系,北京100083 [3]河北医科大学口腔医院口腔颌面外科,河北石家庄050017

出　　处：《华西口腔医学杂志》2007年第1期22-25,28,共5页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目(30271422);国家人事部留学回国人员基金资助项目(20029901);河北省自然科学基金资助项目(2004000624)

摘　　要：目的检测DNA polβ启动子在涎腺腺样囊性癌细胞中的活性,探讨DNA polβ启动子对外源性野生型p53基因表达的影响。方法荧光素酶法测定DNA polβ启动子在涎腺腺样囊性癌细胞中的活性。构建携带人野生型p53基因的真核表达载体,以脂质体法转染涎腺腺样囊性癌SACC-83细胞,逆转录聚合酶链反应(RT-PCR)检测p53基因mRNA的表达。博莱霉素、H2O2及紫外线刺激转染细胞,采用RT-PCR及Western blot法比较DNA损伤下DNA polβ启动子和CMV启动子调控的p53基因及P53蛋白的表达。结果荧光素酶活性分析显示,涎腺腺样囊性癌细胞中DNApolβ启动子活性增高。p53基因的导入使其在涎腺腺样囊性癌细胞中表达增强,DNA polβ启动子组较CMV启动子组更为明显。DNA损伤后,DNA polβ启动子组的p53基因和P53蛋白的表达较CMV启动子组增强(P<0.05)。结论在涎腺腺样囊性癌SACC-83细胞中,DNA polβ启动子的活性增高,DNA polβ启动子能够增强外源性野生型p53基因在涎腺腺样囊性癌细胞中的表达。Objective To evaluate the activity of DNA polβ promoter on p53 gene in salivary adenoid cystic carcinoma（SACC） cells. Methods The luciferase activity was examined and used to evaluate the activity of DNA polβ promoter on SACC-83 cells. Eukaryofic expression plasmids of p53 gene were constructed and stably transfected into SACC-83 Cells. RT-PCR was used to assess the expression of p53 gene. The SACC-83 cells were subjected to the treatments of H202, ultraviolet radiation, Bleocin, and affected p53 mRNA and protein level in SACC-83 cells were charaoterized with RT-PCR and Western blotting. Results The result of luciferase activity proved that the activity of DNA polβ promoter in SACC-83 cells was much higher than that of CMV promoter. The results of RT-PCR suggested that p53 gene with different promoters were all expressed effectively, but the expression efficiency was different. It was greater in DNA polβ group than in CMV group. After DNA damage, p53 gene expression increased and DNA polβ promoter could enhance the expression of p53 gene more than CMV promoter. The results of Western blotting indicated that the expression of P53 protein between the two groups did not show any difference. Conclusion In SACC ceils, the activity of DNA polβ promoter was increased and DNA polβ promoter could enhance the expression of p53.

关 键 词：涎腺腺样囊性癌 DNA polβ P53基因 DNA损伤 基因转染 

分 类 号：R739.8[医药卫生—肿瘤]