反义寡核苷酸作用后肝癌细胞端粒酶活性的变化及差异蛋白的表达  

Activity changes of telomerase and protein expression of hepatocarcinoma cell line SMMC-7721 treated by antisense oligodeoxynucleotide

作　　者：何敏[1] 韦霄[2] 覃健[2] 朱淼[1] 周凌云[2] 黄立新[2] 张志勇[2] 

机构地区：[1]广西医科大学医学科学实验中心,广西南宁530021 [2]广西医科大学公共卫生学院环境卫生学教研室,广西南宁530021

出　　处：《疾病控制杂志》2005年第6期541-544,共4页Chinese Journal of Disease Control and Prevention

基　　金：国家自然科学基金项目(30460121);广西自然科学基金项目(桂科青0135008)

摘　　要：目的利用针对端粒酶RNA的反义寡核苷酸(ASODN)和正义寡核苷酸(NODN)作用于人肝癌细胞SMMC-7721,比较ASODN和NODN作用后癌细胞端粒酶活性及相关蛋白的变化。方法利用实时荧光定量端粒重复序列扩增法(FQ-TRAP法),检测ASODN和NODN作用后肿瘤细胞端粒酶活性变化;蛋白质芯片-飞行时间质谱仪,检测ASODN和NODN作用后特异蛋白质的变化。对照组为未加任何处理因素的SMMC-7721细胞。结果FQ-TRAP法检测CT值,A-SODN组为35.2±2.3,NODN组为26.1±3.5,对照组为18.2±0.7。蛋白质芯片-飞行时间质谱仪检测发现,ASODN组有19个差异蛋白分子高表达,13个差异蛋白分子低表达。NODN组有20个差异蛋白分子高表达,12个差异蛋白分子低表达。所有差异蛋白的分子量分布在3 000 D^10 000 D之间。结论ASODN和NODN均能抑制肝癌细胞端粒酶活性,两者作用SMMC-7721细胞后所表达的差异蛋白十分相似。Objective To investigate the changes of telomerase activity and protein expression of hepatocarcinoma cell line SMMC-7721 treated by antisense oligodexoynuclectide （ASODN） and sense oligodexoynuclectide （NODN）, which aimed at human telomerase RNA. Methods Fluorescent quantitative TRAP（FQ-TRAP） assay and surface ehanced laser desorption time of flight-mass spectrum （SEL- DI-TOF-MS） were used to detect changes of telomerase activity and proteins. The control group was hepatocarcinoma SMMC-7721 cell without any disposal, Results The CT of ASODN, NODN and control group detected by FQ-TRAP assay were 35.2±2.3,26.1±3.5 and 18.2± 0.7 respectively, The results of SELDI-TOF-MS with protein chip assay showed that 19 proteins up-regulated and 13 proteins down-regulated in ASODN group, 20 proteins up-regulated and 12 proteins down-regulated in NODN group. The molecular weight of the these distinct proteins were distributed from 3 000 D to 10 000 D. Conclosions The telomerase activity of hepatocarcinoma cell line SMMC-7721 could he obviously inhibited by ASODN and NODN. The expression of distinct proteins in ASODN group and in NODN group were extremely similar.

关 键 词：寡核苷酸类 反义 肝肿瘤 端粒 未端转移酶 

分 类 号：R341.31[医药卫生—基础医学] R342.4