沉默人乙酰转移酶样蛋白基因对结直肠癌CL187细胞放射敏感性的影响  

Effect of silencing human acetytransferase-like protein on regulation of radiosensitivity in colorectal cancer CL187 cells

作　　者：胡乐林[1,2] 陶慧慧[3] 杨阳丽 陶欣荣[3] 唐小龙[3] HU Lelin;TAO Huihui;YANG Yangli;TAO Xinrong;TANG Xiaolong(First Clinical Hospital,School of Medicine,Anhui University of Science and Technology,Huainan 232001,China;Department of Radiation Oncology,The First Affiliate Hospital of Anhui University of Science and Technology,Huainan 232001,China;Department of Cell Biology,School of Medicine,Anhui University of Science and Technology,Huainan 232001,China)

机构地区：[1]安徽理工大学医学院第一临床学院,淮南232001 [2]安徽理工大学第一附属医院肿瘤放疗科,淮南232001 [3]安徽理工大学医学院生物化学与细胞生物学教研室,淮南232001

出　　处：《辐射研究与辐射工艺学报》2020年第1期23-31,共9页Journal of Radiation Research and Radiation Processing

基　　金：安徽省教育厅自然科学研究项目(KJ2018A0096、KJ2018ZD011);安徽理工大学引进人才科研启动基金和国家大学生创新创业训练计划项目(201910361116)资助~~

摘　　要：探讨沉默人乙酰转移酶样蛋白(Human acetytransferase-like protein,hALP)基因表达对结直肠癌CL187细胞放射敏感性的影响。通过临床数据库挖掘hALP基因在结肠癌组织的表达情况;CL187细胞沉默hALP后接受递增剂量(0 Gy、2 Gy、4 Gy、6 Gy、8 Gy)射线照射,运用克隆增殖实验和细胞存活率检测实验分析细胞放射敏感性变化;结直肠癌CL187细胞沉默hALP后接受8 Gy射线照射,使用流式细胞分析术检测细胞凋亡情况;CL187细胞沉默hALP基因后接受8 Gy射线照射,使用Western blotting检测hALP基因、P53和BAX的蛋白表达水平。结果显示:CL187细胞沉默hALP基因后接受2 Gy射线照射,沉默hALP基因组与对照组的细胞存活分数分别是0.63、0.5,相对生物学效应1.26;沉默hALP基因后射线照射引起的细胞凋亡从19.53%增高到36.49%(p<0.05);沉默hALP基因后射线照射引起P53、BAX表达水平进一步增高。结果提示,沉默hALP基因表达提高了结直肠癌CL187细胞的放射敏感性。hALP基因的表达水平与细胞放射敏感性之间存在相关性,有可能是预测结直肠癌放射敏感性的分子靶标。To investigate the effect of human acetytransferase-like protein(hALP)SiRNA on radiosensitivity of colorectal cancer CL187 cells.For this,the expression of hALP in colon cancer tissues was investigated,based on information available in clinical databases.After hALP knockdown,CL187 cells were irradiated with gradually increasing doses(0 Gy,2 Gy,4 Gy,6 Gy and 8 Gy),and clonal proliferation and cell viability assays were used to detect radiosensitivity.After with the hALP SiRNA duplex,the percentage of cells in which apoptosis was induced by radiation at the dose of 8 Gy was detected by flow cytometric analysis,and the expression of hALP,P53,and BAX was examined by western blotting.Following irradiation,the ratios of surviving fraction between hALP SiRNA and negative control SiRNA groups were 0.63,0.5,respectively,for total doses of 2 Gy(RBE=1.26).Silencing hALP enhanced the radiosensitivity of CL187 cells.Moreover,hALP knockdown increased the percentage of CL187 cells undergoing apoptosis from 19.53%to 36.49%(p<0.05).The expression of hALP,P53 and BAX induced by radiation further increased after hALP knockdown expression.Further,hALP knockdown expression increased radiosensitivity of CL187 cells.The expression of hALP associated with the radiosensitivity of colorectal cancer cells,which is beneficial to screen potential molecular targets and to provide indispensable experimental data for improving the efficacy of radiotherapy.

关 键 词：基因沉默 人乙酰转移酶样蛋白(hALP) 结直肠癌 放射敏感性 

分 类 号：R735.3[医药卫生—肿瘤]