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Myeloid zinc finger 1 mediates sulindac sulfide-induced upregulation of death receptor 5 of human colon cancer cells.

Horinaka M, Yoshida T, Tomosugi M, Yasuda S, Sowa Y, Sakai T - Sci Rep (2014)

Bottom Line: MZF1 directly bound to the putative MZF1-binding site of the DR5 promoter and the binding was increased by sulindac sulfide.The expression of MZF1 was also increased by sulindac sulfide, and MZF1 siRNA attenuated the upregulation of DR5 by sulindac sulfide.These results indicate that sulindac sulfide induces the expression of DR5 by up-regulating MZF1.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Molecular-Targeting Cancer Prevention, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan [2].

ABSTRACT
A combined therapy of sulindac sulfide and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising strategy for the treatment of cancer. Sulindac sulfide had been shown to induce the expression of death receptor 5 (DR5), a receptor for TRAIL, and sensitize cancer cells to TRAIL-induced apoptosis; however, the molecular mechanism underlying the upregulation of DR5 has not yet been elucidated. We demonstrate here that myeloid zinc finger 1 (MZF1) mediates the induction of DR5 by sulindac sulfide. Sulindac sulfide induced the expression of DR5 at the protein and mRNA levels in colon cancer SW480 cells. Furthermore, sulindac sulfide increased DR5 promoter activity. We showed that sulindac sulfide stimulated DR5 promoter activity via the -301 to -253 region. This region contained a putative MZF1-binding site. Site-directed mutations in the site abrogated the enhancement in DR5 promoter activity by sulindac sulfide. MZF1 directly bound to the putative MZF1-binding site of the DR5 promoter and the binding was increased by sulindac sulfide. The expression of MZF1 was also increased by sulindac sulfide, and MZF1 siRNA attenuated the upregulation of DR5 by sulindac sulfide. These results indicate that sulindac sulfide induces the expression of DR5 by up-regulating MZF1.

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Related in: MedlinePlus

Analysis of sulindac sulfide-responsive elements in the DR5 promoter.(a)(b) The luciferase assay was performed as described in Figure 3 with the indicated reporter plasmids. Data represent the means of triplicate experiments (bars, S.D.). +: treated with 200 μM sulindac sulfide for 24 h, −: treated with solvent DMSO. *: p < 0.05.
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f4: Analysis of sulindac sulfide-responsive elements in the DR5 promoter.(a)(b) The luciferase assay was performed as described in Figure 3 with the indicated reporter plasmids. Data represent the means of triplicate experiments (bars, S.D.). +: treated with 200 μM sulindac sulfide for 24 h, −: treated with solvent DMSO. *: p < 0.05.

Mentions: We investigated transcription factors contributing to the upregulation of DR5 by sulindac sulfide. Using a series of deletion mutants in the DR5 promoter flanking the luciferase reporter gene, we performed luciferase assays with or without sulindac sulfide (Fig. 4). The DR5 promoter-luciferase construct pDR5/−448 as well as pDR5PF increased promoter activity by sulindac sulfide more than 2-fold, whereas the promoter activity of pDR5/−198 was not increased (Fig. 4a). This result indicates that sulindac sulfide-responsive elements are located between −448 and −199. Thus, we generated additional reporter plasmids between −448 and −199 and performed luciferase assays (Fig. 4b). Sulindac sulfide enhanced the promoter activity of pDR5/−301 more than 2-fold. On the other hand, pDR5/−252 did not respond to the sulindac sulfide treatment, which indicated that the sulindac sulfide-responsive elements of the DR5 promoter are located within a 50-bp region between −301 and −253 relative to the first base of the translation initiation codon.


Myeloid zinc finger 1 mediates sulindac sulfide-induced upregulation of death receptor 5 of human colon cancer cells.

Horinaka M, Yoshida T, Tomosugi M, Yasuda S, Sowa Y, Sakai T - Sci Rep (2014)

Analysis of sulindac sulfide-responsive elements in the DR5 promoter.(a)(b) The luciferase assay was performed as described in Figure 3 with the indicated reporter plasmids. Data represent the means of triplicate experiments (bars, S.D.). +: treated with 200 μM sulindac sulfide for 24 h, −: treated with solvent DMSO. *: p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126006&req=5

f4: Analysis of sulindac sulfide-responsive elements in the DR5 promoter.(a)(b) The luciferase assay was performed as described in Figure 3 with the indicated reporter plasmids. Data represent the means of triplicate experiments (bars, S.D.). +: treated with 200 μM sulindac sulfide for 24 h, −: treated with solvent DMSO. *: p < 0.05.
Mentions: We investigated transcription factors contributing to the upregulation of DR5 by sulindac sulfide. Using a series of deletion mutants in the DR5 promoter flanking the luciferase reporter gene, we performed luciferase assays with or without sulindac sulfide (Fig. 4). The DR5 promoter-luciferase construct pDR5/−448 as well as pDR5PF increased promoter activity by sulindac sulfide more than 2-fold, whereas the promoter activity of pDR5/−198 was not increased (Fig. 4a). This result indicates that sulindac sulfide-responsive elements are located between −448 and −199. Thus, we generated additional reporter plasmids between −448 and −199 and performed luciferase assays (Fig. 4b). Sulindac sulfide enhanced the promoter activity of pDR5/−301 more than 2-fold. On the other hand, pDR5/−252 did not respond to the sulindac sulfide treatment, which indicated that the sulindac sulfide-responsive elements of the DR5 promoter are located within a 50-bp region between −301 and −253 relative to the first base of the translation initiation codon.

Bottom Line: MZF1 directly bound to the putative MZF1-binding site of the DR5 promoter and the binding was increased by sulindac sulfide.The expression of MZF1 was also increased by sulindac sulfide, and MZF1 siRNA attenuated the upregulation of DR5 by sulindac sulfide.These results indicate that sulindac sulfide induces the expression of DR5 by up-regulating MZF1.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Molecular-Targeting Cancer Prevention, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan [2].

ABSTRACT
A combined therapy of sulindac sulfide and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising strategy for the treatment of cancer. Sulindac sulfide had been shown to induce the expression of death receptor 5 (DR5), a receptor for TRAIL, and sensitize cancer cells to TRAIL-induced apoptosis; however, the molecular mechanism underlying the upregulation of DR5 has not yet been elucidated. We demonstrate here that myeloid zinc finger 1 (MZF1) mediates the induction of DR5 by sulindac sulfide. Sulindac sulfide induced the expression of DR5 at the protein and mRNA levels in colon cancer SW480 cells. Furthermore, sulindac sulfide increased DR5 promoter activity. We showed that sulindac sulfide stimulated DR5 promoter activity via the -301 to -253 region. This region contained a putative MZF1-binding site. Site-directed mutations in the site abrogated the enhancement in DR5 promoter activity by sulindac sulfide. MZF1 directly bound to the putative MZF1-binding site of the DR5 promoter and the binding was increased by sulindac sulfide. The expression of MZF1 was also increased by sulindac sulfide, and MZF1 siRNA attenuated the upregulation of DR5 by sulindac sulfide. These results indicate that sulindac sulfide induces the expression of DR5 by up-regulating MZF1.

Show MeSH
Related in: MedlinePlus