Limits...
Sal-like protein 2 upregulates p16 expression through a proximal promoter element.

Wu Z, Cheng K, Shi L, Li Z, Negi H, Gao G, Kamle S, Li D - Cancer Sci. (2015)

Bottom Line: Promoter-reporter assays of p16(INK4A) and several other tumor-related genes indicated that the Sall2 regulation of these promoters was not significantly different between the two major forms of Sall2 with alternative exon 1 or exon 1A.Finally, to confirm the significance of Sall2-activated p16 expression in cell cycle regulation, we co-transfected the SKOV3 cells with a Sall2 expression construct and a p16 minigene and also co-transfected the ES-2 cells with a Sall2 expression construct and the siRNA against p16 for flow cytometry analysis.Our results showed that Sall2 enhanced the p16 minigene blocking of cell cycle progression and p16 knockdown with siRNA abolished most of the Sall2 inhibition of cell cycle progression.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.

Show MeSH

Related in: MedlinePlus

Identification of a novel Sall2-responsive element on p16 proximal promoter. (a) Luciferase assay with various deletions of p16-luc that covers −1318 to −52 bp promoter region in SKOV3 cells. Sall2 was transiently co-transfected with p16 promoter deletion luciferase constructs into SKOV3 cells with TK-Renilla as an internal control. Luciferase activity was normalized and expressed as fold differences compared with p16-luc. (b) Luciferase assay with various deletions of p16-luc that covers −160 to −52 bp promoter region in SKOV3 cells. A potential Sall2 transcription factor-binding site of p16 promoter sequence is GGGTGGG (−72 to −66 bp). Data is shown as mean ± SD (n = 4; *P < 0.05, t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4376433&req=5

fig04: Identification of a novel Sall2-responsive element on p16 proximal promoter. (a) Luciferase assay with various deletions of p16-luc that covers −1318 to −52 bp promoter region in SKOV3 cells. Sall2 was transiently co-transfected with p16 promoter deletion luciferase constructs into SKOV3 cells with TK-Renilla as an internal control. Luciferase activity was normalized and expressed as fold differences compared with p16-luc. (b) Luciferase assay with various deletions of p16-luc that covers −160 to −52 bp promoter region in SKOV3 cells. A potential Sall2 transcription factor-binding site of p16 promoter sequence is GGGTGGG (−72 to −66 bp). Data is shown as mean ± SD (n = 4; *P < 0.05, t-test).

Mentions: In order to find the Sall2 responsive binding site on p16 promoter, we constructed a series of p16 promoter mutants combining large segment deletions with specific deletions of some of the known transcription factor binding sites and analyzed the reporter luciferase activities in SKOV3 cells (Fig.4).


Sal-like protein 2 upregulates p16 expression through a proximal promoter element.

Wu Z, Cheng K, Shi L, Li Z, Negi H, Gao G, Kamle S, Li D - Cancer Sci. (2015)

Identification of a novel Sall2-responsive element on p16 proximal promoter. (a) Luciferase assay with various deletions of p16-luc that covers −1318 to −52 bp promoter region in SKOV3 cells. Sall2 was transiently co-transfected with p16 promoter deletion luciferase constructs into SKOV3 cells with TK-Renilla as an internal control. Luciferase activity was normalized and expressed as fold differences compared with p16-luc. (b) Luciferase assay with various deletions of p16-luc that covers −160 to −52 bp promoter region in SKOV3 cells. A potential Sall2 transcription factor-binding site of p16 promoter sequence is GGGTGGG (−72 to −66 bp). Data is shown as mean ± SD (n = 4; *P < 0.05, t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4376433&req=5

fig04: Identification of a novel Sall2-responsive element on p16 proximal promoter. (a) Luciferase assay with various deletions of p16-luc that covers −1318 to −52 bp promoter region in SKOV3 cells. Sall2 was transiently co-transfected with p16 promoter deletion luciferase constructs into SKOV3 cells with TK-Renilla as an internal control. Luciferase activity was normalized and expressed as fold differences compared with p16-luc. (b) Luciferase assay with various deletions of p16-luc that covers −160 to −52 bp promoter region in SKOV3 cells. A potential Sall2 transcription factor-binding site of p16 promoter sequence is GGGTGGG (−72 to −66 bp). Data is shown as mean ± SD (n = 4; *P < 0.05, t-test).
Mentions: In order to find the Sall2 responsive binding site on p16 promoter, we constructed a series of p16 promoter mutants combining large segment deletions with specific deletions of some of the known transcription factor binding sites and analyzed the reporter luciferase activities in SKOV3 cells (Fig.4).

Bottom Line: Promoter-reporter assays of p16(INK4A) and several other tumor-related genes indicated that the Sall2 regulation of these promoters was not significantly different between the two major forms of Sall2 with alternative exon 1 or exon 1A.Finally, to confirm the significance of Sall2-activated p16 expression in cell cycle regulation, we co-transfected the SKOV3 cells with a Sall2 expression construct and a p16 minigene and also co-transfected the ES-2 cells with a Sall2 expression construct and the siRNA against p16 for flow cytometry analysis.Our results showed that Sall2 enhanced the p16 minigene blocking of cell cycle progression and p16 knockdown with siRNA abolished most of the Sall2 inhibition of cell cycle progression.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai, China.

Show MeSH
Related in: MedlinePlus