Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells.
Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells.
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DOI：
10.1186/1471-2199-12-27
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发表时间：
2011-06-10
影响因子：
--
通讯作者：
Krawczyk Z
中科院分区：
文献类型：
--
作者：
Fiszer-Kierzkowska A;Vydra N;Wysocka-Wycisk A;Kronekova Z;Jarząb M;Lisowska KM;Krawczyk Z
在对大鼠应激诱导的Hspa 1b（hsp70.1）基因的功能研究中，我们注意到一些用于转染的基于脂质体的DNA载体诱导其启动子活性。该观察结果涉及市售脂质体制剂（LA）、Lipofectin和Lipofectamine 2000。这项工作的目的是更好地了解这一现象的机制及其潜在的生物和实际后果。我们发现，Hspa 1b启动子驱动的报告基因激活的情况下，瞬时转染和稳定转染细胞用LA处理。使用几个含有不同片段的Hspa 1b启动子的缺失克隆，我们发现负责最有效的LA驱动的诱导的调控元件位于相对于转录起始位点的核苷酸-269和+85之间。进一步的研究表明，诱导机制是独立的经典的HSE-HSF相互作用，是负责在热应激过程中的基因激活。使用DNA微阵列，我们还检测到显着激活的内源性Hspa 1b基因与Lipofectamine 2000处理的细胞。其他几个应激基因也被诱导，沿着许多参与细胞代谢、细胞周期控制和促凋亡途径的基因。我们的观察结果表明，i）一些阳离子脂质体可能不适合热休克蛋白启动子的功能研究，ii）脂质体转染可能会导致非预期的变化，在整体基因表达的转染细胞。
During functional studies on the rat stress-inducible Hspa1b (hsp70.1) gene we noticed that some liposome-based DNA carriers, which are used for transfection, induce its promoter activity. This observation concerned commercial liposome formulations (LA), Lipofectin and Lipofectamine 2000. This work was aimed to understand better the mechanism of this phenomenon and its potential biological and practical consequences. We found that a reporter gene driven by Hspa1b promoter is activated both in the case of transient transfections and in the stably transfected cells treated with LA. Using several deletion clones containing different fragments of Hspa1b promoter, we found that the regulatory elements responsible for most efficient LA-driven inducibility were located between nucleotides -269 and +85, relative to the transcription start site. Further studies showed that the induction mechanism was independent of the classical HSE-HSF interaction that is responsible for gene activation during heat stress. Using DNA microarrays we also detected significant activation of the endogenous Hspa1b gene in cells treated with Lipofectamine 2000. Several other stress genes were also induced, along with numerous genes involved in cellular metabolism, cell cycle control and pro-apoptotic pathways. Our observations suggest that i) some cationic liposomes may not be suitable for functional studies on hsp promoters, ii) lipofection may cause unintended changes in global gene expression in the transfected cells.
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影响因子：
2.5
作者：
BRAZOLOT, CL;PETITTE, JN;GIBBINS, AMV
通讯作者：
GIBBINS, AMV
DOI：
10.1073/pnas.0702557104
发表时间：
2007-05-08
影响因子：
11.1
作者：
Nagy, Eniko;Balogi, Zsolt;Vigh, Laszlo
通讯作者：
Vigh, Laszlo
影响因子：
6.4
作者：
Sochanik, A;Kaida, I;Szala, S
通讯作者：
Szala, S
DOI：
10.1196/annals.1391.005
发表时间：
2007-01-01
期刊：
STRESS RESPONSES IN BIOLOGY AND MEDICINE
影响因子：
--
作者：
Akerfelt, Malin;Trouillet, Diane;Sistonen, Lea
通讯作者：
Sistonen, Lea
影响因子：
3.1
作者：
POTHIER, F;OUELLET, M;GUERIN, SL
通讯作者：
GUERIN, SL