Change of Ankyrin G Promoter Activity Following Treatment with Sodium Valproate at Different Concentrations.
- Author:
Cui LIU
1
;
Jie WU
1
;
Xiao-long SUI
1
;
Yan-hong LI
1
;
Yun-lin HAN
1
;
Yu-huan XU
1
;
Lan HUANG
1
;
Hua ZHU
1
;
Shu-li SHENG
1
;
Chuan QIN
1
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Ankyrins; Cell Line; Genetic Vectors; Humans; Luciferases; Mice; Mice, Inbred C57BL; Promoter Regions, Genetic; Up-Regulation; Valproic Acid
- From: Acta Academiae Medicinae Sinicae 2015;37(5):508-513
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate whether sodium valproate (VPA) directly regulates the activity of Ankyrin G(AnkG) promoter in vitro.
METHODSThe mouse AnkG promoter sequence was identified by comparing both human and mouse AnkG promoter sequences. The promoter was amplified from C57BL/6 mouse genome DNA and cloned into pGL3 Luciferase reporter vector. The Luciferase activity was detected in N2a and 293T cells and then treated with 0,0.5, and 1 mmol/L VPA for 12 h. The transcription activity of AnkG promoter in cells and the activity of VPA-treated Luciferase reporter vector in cells were detected using dual Luciferase reporter assay.
RESULTSThe AnkG promoter clone and its expression vector were successfully established, as confirmed by enzyme digestion and sequencing. The AnkG promoter showed high transcription activity in both N2a and 293T cells. The Luciferase activity was significantly induced following 0.5 mmol/L VPA treatment in both N2a and 293T cells. CONCLUSIONS VPA can up-regulate the AnkG expression via directly increasing its transcription activity. Thus, the in vivo AnkG expression may be directly regulated by the VPA at transcriptional level.