沉默UVRAG基因在二烯丙基二硫诱导K562细胞中caspase3的表达  

The Expression of Caspase3 in Leukemia K562 Cell Through silence UVRAG induced Diallyl Disulfide

作　　者：肖艳芳[1] 殷小成[1] 彭艳辉[1] 杨云华[1] 罗永姣[1] 

机构地区：[1]南华大学附属第一医院儿科,湖南衡阳421001

出　　处：《现代生物医学进展》2015年第22期4268-4271,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31271482)

摘　　要：目的:沉默UVRAG基因在DADS诱导K562细胞中观察caspase3的表达。方法:以K562细胞为细胞模型,将构建成功并筛选出最有效干扰抑制UVRAG基因的si RNA序列片段采用lipofectamine TM2000脂质体转染法转染白血病K562细胞,组别为:空白对照组,转染试剂组、阴性对照组,阳性对照组,转染24小时后,利用QT-PCR检测UVRAG m RNA的表达水平以此观察干扰效果,再以40 mg/LDADS处理转染试剂组12小时,采用蛋白印迹(Western Blotting)技术检测凋亡相关基因caspase3的表达。结果:QT-PCR显示:与空白组相比,UVRAG m RNA表达明显减少,表明沉默UVRAG基因成功;Western Blotting显示:DADS处理的干扰成功的K562细胞12小时后,检测到caspase3的蛋白表达水平下降。结论:沉默UVRAG基因能使白血病K562细胞中凋亡相关基因caspase3的蛋白表达水平下降,提示抑制UVRAG表达的同时也可能抑制DADS诱导K562细胞的凋亡。Objective： DADS induced the K562 cells in order to observe the expession of caspase3 through silencing UVRAG genes. Methods： The study choose K562 cells for cell model,successfully constructed and selected the most effective gene sequence fragments of siRNA about the gene of UVRAG tranfect to the K562cells using lipofectamine TM2000-1iposomal. There are divided into control group,transfection reagent group,negative control group,positive control group, The interference effection is observed by the QT-PCR detecting the expression levels of UVRAG mRNA after 24 hours transfection, then using 40 mg/LDADS effects the group of transfection 12 hours. Western blotting is used to detect the expression of caspase3 protein induced K562 cell apoptosis. Results： compared with the blank group, the UVRAG mRNA expression levels were lower ,it shows the silence of UVRAG successfully, transferred siRNA-UVRAG of K562 cells in 24 hours,the protein levels of caspase3 is decreased. Conclusions： The protein expression of apoptosis-related genes caspase3 is declined through silencing the UVRAG genes in leukemia K562 cells,it confirmed that inhibition the expression of UVRAG may inhhibit the occurrence of apoptosis ofK562 cells.

关 键 词：二烯丙基二硫 K562细胞 凋亡 抗紫外线相关基因 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R33[医药卫生—人体生理学]