亨廷顿蛋白相关蛋白1基因沉默对小鼠胰岛β细胞系NIT细胞凋亡的影响  

Effect of silencing Huntingtin-associated protein 1 on the apoptosis of mouse islet beta cells line NIT cells

作　　者：陈世新[1] 张婵[1] 侯杰[1] 任艳华[1] 廖敏[1] 

机构地区：[1]温州医学院人体解剖学与组织胚胎学系,浙江温州325035

出　　处：《解剖学报》2010年第3期379-382,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30671106);浙江省自然科学基金资助项目(20061832);浙江省教育厅资助项目(20070913)

摘　　要：目的探讨沉默亨廷顿蛋白相关蛋白1(HAP1)基因表达对小鼠胰岛β细胞株-NIT细胞凋亡的影响。方法化学合成针对小鼠HAP1基因的siRNA,转染NIT细胞,观察干扰效果;通过膜联蛋白V/碘化丙啶(AnnexinⅤ/PI)染色和原位末端核苷酸标记法(TUNEL),检测沉默HAP1表达后NIT细胞的凋亡;通过AnnexinⅤ/PI染色检测沉默HAP1表达后,链脲佐菌素(STZ)所诱导的NIT细胞凋亡。结果靶向HAP1的siRNA能有效抑制NIT细胞HAP1的表达;HAP1 siRNA实验组,NIT细胞凋亡数增多,凋亡率显著高于空白对照组(P<0.01);STZ可明显诱导NIT细胞的凋亡,沉默HAP1的表达能增加STZ所诱导的NIT细胞凋亡。结论沉默HAP1的表达可以增加小鼠胰岛β细胞株NIT细胞的凋亡,同时也能促进凋亡诱导剂(STZ)所诱导的胰岛NIT细胞的凋亡。Objective To investigate the effect of silencing Huntingtin-associated protein 1 （ HAP1 ） on apoptosis of mouse paneraetie islet beta cells line（ NIT cells）. Methods 1. The NIT cells were transfeeted with chemically synthesized siRNA targeting HAP1 for comprehensive evaluation of its silence efficacy. 2. The expression of the silencing HAP1 was detected by Annexin V and PI, and TdT-mediated dUTP-biotin nick end-labeling （TUNEL） to evaluate its efficacy on the apoptosis of NIT cells. 3. The expression of the silencing HAP1 was detected by Annexin V and PI to evaluate its efficacy on the streptozotocin （STZ）-induced apoptosis of NIT cells. Results The expression of silencing HAP1 in NIT cells was inhibited significantly by chemically synthesized siRNA targeting HAP1 ; The number of the apoptosis of NIT cells based on estimates of the difference between experimental and control group, it was markedly increased in the group transfected with chemically synthesized siRNA targeting HAP1 as compared with the control group （P 〈 0. 01 ） ; STZ-induced apoptosis of NIT cells was significantly increased （P 〈 0.01 ） , and the expression of silencing HAP1 was sufficient to promote STZ-induced apoptosis of NIT cells （P 〈 0. 01 ）. Conclusion The expression of silencing HAP1 increases the apoptosis of mice pancreatic islet beta cells line NIT, and it enhances the capability of apoptosis-inducer, STZ and to quicken the apoptosis of NIT cells in simultaneity.

关 键 词：亨廷顿蛋白相关蛋白1 NIT细胞 SIRNA 膜联蛋白V/碘化丙啶涂色 原位末端核苷酸标记法 小鼠 

分 类 号：R322.57[医药卫生—人体解剖和组织胚胎学]