腺病毒携带shFOXM1对ACC-2细胞作用机制的研究  

Research on the Mechanism of ACC-2 Cell Regulation by Adenovirus Harboring shFOXM1

作　　者：仇静[1] 张月[1] 陈书兰[1] 刘岚[1] 刘峤峤 张广耘[1] 袁晓[1] 

机构地区：[1]青岛市市立医院,山东青岛266000

出　　处：《现代生物医学进展》2016年第7期1249-1252,1324,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31170891)

摘　　要：目的:研究携带特异性抑制叉头框蛋白M1(Forkhead box protein M1,FOXM1)表达的腺病毒(Ad5.sh FOXM1)对人涎腺腺样囊性癌(Adenoidcystic carcinoma,ACC)细胞周期的调控、细胞凋亡的影响及其作用机理。方法:根据人FOXM1 m RNA的序列,设计合成针对FOXM1基因的三对sh RNA,转染ACC-2细胞系,利用实时荧光定量PCR、Western blot筛选获得最佳干扰片段,并构建入腺病毒载体,利用流式细胞术、MTT、Western blot等检测其对ACC-2细胞周期、细胞凋亡的调控及分子作用。结果:1成功筛选获得了抑制FOXM1表达的sh RNA,并构建了特异性抑制ACC-2细胞中FOXM1表达的腺病毒Ad5-sh FOXM1;2Ad5-sh FOXM1能明显抑制ACC细胞的增殖,并引起ACC-2细胞S期的阻滞(P<0.05);2Ad5-sh FOXM1通过下调c-Myc蛋白的表达抑制了细胞的增殖,通过下调P21Cip1,P27Kip1蛋白的表达抑制了ACC-2细胞周期(P<0.05)。结论:下调FOXM1的表达能够明显抑制ACC-2细胞周期和ACC-2细胞的增殖。Objective： Research on the specific inhibition of forkhead box protein M1（FOXM1） expression by adenovirus（Ad5.shFOXM1） and the mechanism of cell cycle and cell apoptosis regulation in human adenoidcystic carcinoma（ACC）. Methods： According to the sequence of human FOXM1 m RNA, we designed and synthesis three FOXM1 sh RNA, which was transfected into ACC-2 cells.We identified the most potently suppressed FOXM1 m RNA by Real-time PCR and Western blot. We constructed adenovirus vector harboring the best interference fragment, and detected the cell cycle and cell apoptosis regulation and mechanism by flow cytometry, MTT,Western blot in ACC-2. Results： We identified the most potent sh RNA targeting FOXM1. After constructing of Ad5.shFOXM1, the Ad5.shFOXM1 can significantly inhibit the proliferation of ACC-2 cells, and caused the block of ACC-2 cells in the S phase（P0.05）;Ad5-shFOXM1 inhibited cell proliferation through down regulating the expression of c-Myc, and down regulating the expression of P21Cip1, P27Kip1 protein expression in ACC-2 cell（P0.05）. Conclusion： Down-regulation of FOXM1 expression can inhibit the cell cycle and cell proliferation of ACC.

关 键 词：叉头框蛋白M1 人涎腺腺样囊性癌 细胞周期 

分 类 号：R-33[医药卫生] Q78[生物学—分子生物学]