Effect of Krüppel-like factor 4 overexpression on heat stress-induced apoptosis of Raw264.7 macrophages.
- Author:
Mei-dong LIU
1
;
Ying LIU
;
Jun-wen LIU
;
Hua-li ZHANG
;
Xian-zhong XIAO
Author Information
1. Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Apoptosis;
genetics;
Cell Line;
Genetic Vectors;
Heat-Shock Response;
Kruppel-Like Transcription Factors;
genetics;
Macrophages;
cytology;
Mice;
Plasmids;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2007;32(6):1002-1006
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To observe the effect of Krüppel-like factor 4 (KLF4) overexpression on heat stress-induced apoptosis of Raw264.7 macrophages.
METHODS:The fragment containing full length mouse KLF4 cDNA coding sequence was inserted into the pcDNA3.1 vector and Raw264.7 macrophages were transfected with pcDNA3.1-KLF4 plasmids using lipofectamine.The expression of KLF4 was examined by Western blot in the Raw264.7 macrophages stably transfected with pcDNA3.1- KLF4 plasmids. Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1-KLF4 were exposed to heat stress (42 degrees C) for 1h and recovered at 37 degrees C for 12h. Flow cytometry, Hoechest 33258 staining assay, and DNA ladder assays were performed to assess the apoptosis.
RESULTS:The KLF4 overexpressed Raw264.7 macrophages were established. After the heat stress, flow cytometry showed that apoptotic cells increased significantly in KLF4 overexpressed cells compared with the vector control; Hoechest 33258 staining was characterized with classical changes including apoptotic body, and nuclear condensation. Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly.
CONCLUSION:KLF4 overexpression can increase heat stress-induced apoptosis of Raw264.7 macrophages.
