[Immortalization of embryonic fibroblasts in heat shock transcription factor 1 knockout mouse].
OBJECTIVE
To establish immortalized embryonic fibroblast lines in heat shock transcription factor 1 (HSF1) HSF1-/- and HSF1+/+ mice and to provide experimental models to study the function of HSF1.


METHODS
A mammalian expression vector (pSV3neo) containing the SV40 large T antigen was used to transfect the HSF1-/- and HSF1+/+ mouse embryonic fibroblast using Lipofectamine 2000. Colonies were screened by G418 and expanded to immortalized cell lines. PCR was used to detect the integration of the large T antigen with genome in the mouse embryonic fibroblast. Expression of SV40 large T antigen gene in expanded cells was identified by RT-PCR. HSP70 expression was examined by Western blot in the embryonic fibroblast lines.


RESULTS
The stable growth and serial propagation were observed in the HSF1-/- and HSF1+/+ cell lines for six months. The mRNA of SV40 T antigen gene expressed in the two cell lines. HSP70 expression could not be induced in the heat-treated HSF1-/- mouse embryo fibroblasts.


CONCLUSIONS
The immortalized cells of HSF1+/+ and HSF1-/- mouse embryo fibroblasts are successfully established.
Liu
Mei-dong
MD
Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, China.
Zhang
Hua-li
HL
Gong
Huan-yu
HY
Chen
Guang-wen
GW
Wang
Kang-kai
KK
Shun-mei
E
E
Xiao
Xian-zhong
XZ
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000494:pharmacology*; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics*; D004622:Embryo, Mammalian; D005260:Female; D005347:Fibroblasts / Q000166:cytology*; D000076249:Heat Shock Transcription Factors; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D014157:Transcription Factors / Q000235:genetics*
D000952:Antigens, Polyomavirus Transforming; D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; C490017:Hsf1 protein, mouse; D014157:Transcription Factors