反义核苷酸沉默Livin基因对喉鳞癌细胞凋亡影响的体外研究  

Influence of antisense oligodeoxynucleotides targeting livin on apoptosis of laryngeal squamous cell carcinoma in vitro

作　　者：冯俊[1] 李丽[2] 杨九梅 彭涛[1] 王朝莉[3] 

机构地区：[1]川北医学院第二临床医学院.南充市中心医院耳鼻咽喉科,四川南充637000 [2]川北医学院病理教研室,四川南充647000 [3]川北医学院分子生物研究所,四川南充637000

出　　处：《西部医学》2016年第9期1207-1210,共4页Medical Journal of West China

基　　金：四川省教育厅基金资助(13ZB0241)

摘　　要：目的为了保护喉功能和提高喉鳞癌病人的生活质量,我们设计反义Livin作用于喉鳞状细胞癌来减少Livin表达,分析在体外对喉鳞状细胞癌细胞凋亡的影响。方法用real-time quantitative PCR对AS-ODNs、SenODNs、Mis-ODNs、hep-2组(Control)的Livin mRNA进行分析;用Western blot对AS、Sen、Mis、与hep-2组的Livin蛋白进行分析;用流式细胞仪分析喉癌细胞的凋亡情况。结果AS-ODNs、Sen-ODNs、Mis-ODNs与Control组的Livin mRNA相对表达量以AS-ODNs组最低,为(24.03±7.02)%,AS组与Sen、Mis、Control组比较,差异均有统计学意义(P<0.05)。AS、Sen、Mis与Control组的Livin蛋白相对表达量以AS-ODNs组最低,为(11.65±3.68)%,AS组与Sen、Mis、Control组比较,差异亦有统计学意义(均P<0.05)。将AS、Sen、Mis转染喉鳞状细胞癌细胞,行流式细胞仪检测,结果发现AS组的凋亡率为(36.54±6.04)%,远高于Sen、Mis、Control组,差异均有统计学意义(P<0.05)。结论在体外,Livin反义多核苷酸降低了喉鳞状细胞癌中Livin的表达,同时增强了喉鳞状细胞癌细胞的凋亡。Objective To conserve laryngeal function and elevate living quality of laryngeal squamous cell carcinoma （LSCC） patients, we designed antisense oligodeoxynucleotides （AS-ODNs） to reduce expression of livin and to learn the apoptosis of hep-2 （Human epidermoid laryngeal carcinoma） cells in vitro. Methods The expression of livin mRNA in hep-2 cells was examined by real-time quantitative PCR. In groups of AS, Sen, Mis and hep-2, the expression of livin protein were examined by western blotting respectively, and the cell apoptosis analyzed by flow cytometry. Results The relative Livin mRNA expression in hep-2 cells treated with Livin AS-ODNs was decreased to （24.03±7.02）% of that in untreated cells respectively （P〈0.05）. The relative livin protein expression in hep-2 cells treated with Livin AS-ODNs were decreased to （11.65±3.68）% of that in untreated cells respectively （P〈0.05）. The apoptotic rate for the group treated with livin AS-ODNs was （36.54± 6.04）%, which was significantly higher than that of others （P〈0.05）. Conclusion AS-ODNs of livin reduces livin gene expression and enhances hep-2 cells apoptosis in vitro.

关 键 词：LIVIN 反义核苷酸 喉鳞状细胞癌 凋亡 

分 类 号：R739.65[医药卫生—肿瘤]