干涉鸡热休克蛋白70基因表达诱导鸡胚成纤维细胞凋亡

畜牧兽医学报

干涉鸡热休克蛋白70基因表达诱导鸡胚成纤维细胞凋亡

杨帆^1,2，汤承¹ ，岳华^1*

（1.西南民族大学，成都 610041； 2. 成都康弘药业集团股份有限公司，成都 610036）

收稿日期:2012-09-20 出版日期:2013-03-23 发布日期:2013-03-25
通讯作者: 岳华，教授， E-mail：yhua900@163.com
作者简介:杨帆(1984-)，男，甘肃徽县人，硕士，主要从事病原分子生物学研究，E-mail：yfqh84@yahoo.com.cn
基金资助:
自然科学基金（31172307）

Interference of Chicken Heat Shock Protein 70 Gene Expression Induced ChickenEmbryo Fibroblast Apoptosis

YANG Fan^1,2, TANG Cheng¹, YUE Hua^1*

(1. Southwest University for Nationalities, Chengdu 610041, China; 2. Chengdu Kanghong Pharmaceutical Group Co., Ltd., Chengdu 610036, China)

Received:2012-09-20 Online:2013-03-23 Published:2013-03-25

摘要/Abstract

摘要：

为研究鸡热休克蛋白70（HSP70）与细胞凋亡的相关性及分子机制，通过构建表达靶向鸡hsp 70的短发夹样结构小干扰RNA (shRNA)质粒载体，转染鸡胚成纤维细胞（CEF），干涉后18、24、28 h用Annexin V-FITC方法检测干涉hsp70表达后细胞凋亡情况，同时用荧光定量RT-PCR测定凋亡相关基因Caspase3和Caspase8 的表达水平。结果显示，与未干涉组相比，干涉组hsp70被分别敲低了75%、83%和96%，凋亡细胞数随着干涉时间的延长而增加，Caspase3表达水平升高，其表达量分别上调至250%、140%和110%， Caspase-8表达水平下降，其表达水平分别下降至76%、80%和34%。由此可见，鸡的hsp 70可能主要通过线粒体途径保护CEF对抗凋亡。

关键词: 热休克蛋白 70, RNA 干涉, 鸡胚成纤维细胞, 凋亡, 半胱氨酸蛋白酶

Abstract:

The purpose of this paper is to study the relationship and molecular mechanisms between chicken heat shock protein 70 (HSP70) and apoptosis. Plasmid vectors expressing short hairpin-like structure small interfering RNA (shRNA) targeting chicken HSP70 were constructed and transfected into the chicken embryo fibroblast (CEF) in this study. At 18, 24, 28 h post interference, the CEF apoptosis was observed by Annexin V-FITC method following interference of hsp70 expression and caspase3 ＆ caspase8 were quantified by real-time RT-PCR. The results showed that, compared with the mock cell, 1) Apoptotic cells increased as the interference time extended. 2) The hsp70 in CEF cells decreased by 75%, 83% and 96% while caspase3 increased to 250%, 140% and 110%, and caspase8 decreased to 76%, 80% and 34% at 18, 24, 28 h post interference respectively. The results projected that chicken hsp70 protects the CEF cells against apoptosis mainly via the mitochondrial pathway.

Key words: heat shock protein 70, RNA interference, chicken embryo fibroblast, apoptosis, caspase

中图分类号:

S852.23

杨帆，汤承，岳华. 干涉鸡热休克蛋白70基因表达诱导鸡胚成纤维细胞凋亡[J]. 畜牧兽医学报.

YANG Fan, TANG Cheng, YUE Hua. Interference of Chicken Heat Shock Protein 70 Gene Expression Induced ChickenEmbryo Fibroblast Apoptosis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA.

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