放射诱导p16基因胰腺癌靶向性表达的研究  

Pancreatic carcinoma targeted expression of p16 gene controlled by irradiation via Egr-1 promoter

作　　者：马红兵[1] 王西京[1] 胡海涛[2] 狄政莉[1] 夏辉[3] 李铮[3] 马洁[3] 白明华[1] 韩志楷[3] 吴丛梅[4] 

机构地区：[1]西安交通大学第二医院肿瘤科,陕西西安710004 [2]西安交通大学医学院解剖学教研室,陕西西安710061 [3]中国医学科学院肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [4]汕头大学医学院生物学教研室,广东汕头515031

出　　处：《西安交通大学学报（医学版）》2005年第1期68-70,共3页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省科技攻关项目资助(No.2002K10 G3);西安交通大学第二医院科研基金(No.2001YJ 04)

摘　　要：目的　构建 pcDNA3.1 Egr.1p-p16重组质粒并检测其在人胰腺癌 JF305细胞中的辐射诱导表达。方法　将人p16 cDNA基因连接到有辐射诱导特性的早期反应因子 Egr.1p的下游,构建成 pcDNA3.1 -Egr.1p- p16 重组质粒,利用脂质体介导转染人胰腺癌细胞系 JF305 细胞;采用 RT- PCR方法和 Western blot方法检测不同剂量 X射线照射后,被转染细胞中 p16 的转录和表达水平。结果　酶切鉴定证实 pcDNA3. 1 Egr. 1p -p16 重组质粒构建正确。被pcDNA3.1 Egr.1p -p16重组质粒转染的人胰腺癌 JF305细胞经不同剂量 X射线照射后,p16 基因表达均高于未照射组。结论　X射线可诱导 pcDNA3.1 -Egr.1p -p16重组质粒在人胰腺癌 JF305细胞中表达增强。Objective To construct a recombinant plasmid pcDNA3.1-Egr.1p-p16 and investigate the expression of p16 induced by irradiation in JF305 cells. Methods Human p16cDNA was ligated to the downstream of Egr-promoter to construct pcDNA3.1-Egr.1p-p16 plasmid. The recombined plasmids were transfected into JF305 cells with liposome. RT-PCR and Western blot were used to detect the dose effect and time course manners of p16 mRNA level and p16 protein expression after irradiation. Results Restrictive enzyme digestion showed pcDNA 3.1-Egr.1p-p16 was correctly constructed. The p16 expression in cells transfected with pcDNA3.1-Egr.1p-p16 induced by different doses of irradiation was higher than that of sham-irradiation group. Conclusion X ray can induce and enhance the expression of pcDNA3.1-Egr.1p-p16 plasmid in JF305 cells.

关 键 词：EGR-1启动子 X射线 P16表达 胰腺癌细胞系 

分 类 号：R146[医药卫生—公共卫生与预防医学]