肖艳芳 殷小成△ 彭艳辉 杨云华 罗永姣.沉默UVRAG 基因在二烯丙基二硫诱导K562细胞中caspase3 的表达[J].现代生物医学进展英文版,2015,15(22):4268-4271. |
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 silenceUVRAG induced Diallyl Disulfide |
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DOI: |
中文关键词: 二烯丙基二硫 细胞 凋亡 抗紫外线相关基因 半胱氨酸天冬氨酸蛋白酶3 |
英文关键词: Diallyl disulfide cell Apoptosis UVRAG Caspase3 |
基金项目:国家自然科学基金项目(31271482) |
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中文摘要: |
目的:沉默UVRAG 基因在DADS诱导细胞中观察caspase3的表达。方法:以细胞为细胞模型,将构建成功并
筛选出最有效干扰抑制UVRAG基因的siRNA序列片段采用lipofectamine TM2000 脂质体转染法转染白血病细胞, 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 mRNA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 mRNA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 诱导细胞的凋亡。 |
英文摘要: |
Objective:DADS induced the cells in order to observe the expession of caspase3 through silencing UVRAG
genes.Methods:The study choose cells for cell model,successfully constructed and selected the most effective gene sequence
fragments of siRNA about the gene of UVRAG tranfect to the cells using lipofectamine TM2000-liposomal. 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 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 cells in 24 hours,the protein levels of caspase3 is decreased.Conclusion:The protein expression of
apoptosis-related genes caspase3 is declined through silencing the UVRAG genes in leukemia cells,it confirmed that inhibition the
expression of UVRAG may inhhibit the occurrence of apoptosis of cells. |
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