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Cooperatively transcriptional and epigenetic regulation of sonic hedgehog overexpression drives malignant potential of breast cancer.

Duan ZH, Wang HC, Zhao DM, Ji XX, Song M, Yang XJ, Cui W - Cancer Sci. (2015)

Bottom Line: Comprehensive understanding of the regulation mechanism of Shh in cancer cells is necessary to find an effective approach to selectively block its tumorigenic function.Moreover, in vitro data demonstrated that both NF-κB activation and hypomethylation in promoter region were positively associated with the overexpression of Shh.Furthermore, the biological function data indicated that overexpressed Shh enhanced the self-renewal capacity and migration ability of breast cancer cells, which could be augmented by promoter demethylation and NF-κB activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, College of Life Science and Biopharmaceutical of Shenyang Pharmaceutical University, Shenyang, China.

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Related in: MedlinePlus

The correlation of nuclear factor-kappa B (NF-κB) expression and promoter methylation status with Sonic hedgehog (Shh) expression in breast cell lines. (a) The expression of Shh and nuclear NF-κB in breast cancer cell lines. (b) The expression of Shh mRNA in breast cancer cell lines. MDA-MB-231 (MM231); MDA-MB-436 (MM436). (c) Methylation status of Shh promoter in breast cell lines and the effect of 5-azacytidine (5-Aza) treatment. MM231 and Bcap37 cells were treated with 20 μM 5-Aza for 72 h. The genomic DNA was extracted for methylation-specific (MS)-PCR. M, methylated; UM, non-methylated. (d) Expression of Shh in breast cell lines after 5-Aza treatment.
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fig02: The correlation of nuclear factor-kappa B (NF-κB) expression and promoter methylation status with Sonic hedgehog (Shh) expression in breast cell lines. (a) The expression of Shh and nuclear NF-κB in breast cancer cell lines. (b) The expression of Shh mRNA in breast cancer cell lines. MDA-MB-231 (MM231); MDA-MB-436 (MM436). (c) Methylation status of Shh promoter in breast cell lines and the effect of 5-azacytidine (5-Aza) treatment. MM231 and Bcap37 cells were treated with 20 μM 5-Aza for 72 h. The genomic DNA was extracted for methylation-specific (MS)-PCR. M, methylated; UM, non-methylated. (d) Expression of Shh in breast cell lines after 5-Aza treatment.

Mentions: To confirm the correlation among NF-κB nuclear expression, promoter methylation status with Shh expression, several breast cell lines, MCF-7, Bcap37, MDA-MB-436 and MDA-MB-231, were used in our experiments. As shown in Figure2a, MCF-7 cells expressed Shh at a relatively higher level, Bcap37 and MDA-MB-231 cells displayed a moderate expression, but MDA-MB-436 cells expressed Shh at a lower level. Interestingly, a similar pattern was shown in NF-κB nuclear expression, suggesting a positive correlation between NF-κB and Shh. As the transcription factor, NF-κB regulates target gene expression in mRNA level. Thus, we also detected the expression of Shh in mRNA level using quantitative RT-PCR. Consistent with protein expression data, Shh was expressed at a higher level in MCF-7 cells, but not in Bcap37, MDA-MB-436 and MDA-MB-231 cells (see Fig.2b), demonstrating that the crucial regulation of Shh expression is presented in the transcription level.


Cooperatively transcriptional and epigenetic regulation of sonic hedgehog overexpression drives malignant potential of breast cancer.

Duan ZH, Wang HC, Zhao DM, Ji XX, Song M, Yang XJ, Cui W - Cancer Sci. (2015)

The correlation of nuclear factor-kappa B (NF-κB) expression and promoter methylation status with Sonic hedgehog (Shh) expression in breast cell lines. (a) The expression of Shh and nuclear NF-κB in breast cancer cell lines. (b) The expression of Shh mRNA in breast cancer cell lines. MDA-MB-231 (MM231); MDA-MB-436 (MM436). (c) Methylation status of Shh promoter in breast cell lines and the effect of 5-azacytidine (5-Aza) treatment. MM231 and Bcap37 cells were treated with 20 μM 5-Aza for 72 h. The genomic DNA was extracted for methylation-specific (MS)-PCR. M, methylated; UM, non-methylated. (d) Expression of Shh in breast cell lines after 5-Aza treatment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556399&req=5

fig02: The correlation of nuclear factor-kappa B (NF-κB) expression and promoter methylation status with Sonic hedgehog (Shh) expression in breast cell lines. (a) The expression of Shh and nuclear NF-κB in breast cancer cell lines. (b) The expression of Shh mRNA in breast cancer cell lines. MDA-MB-231 (MM231); MDA-MB-436 (MM436). (c) Methylation status of Shh promoter in breast cell lines and the effect of 5-azacytidine (5-Aza) treatment. MM231 and Bcap37 cells were treated with 20 μM 5-Aza for 72 h. The genomic DNA was extracted for methylation-specific (MS)-PCR. M, methylated; UM, non-methylated. (d) Expression of Shh in breast cell lines after 5-Aza treatment.
Mentions: To confirm the correlation among NF-κB nuclear expression, promoter methylation status with Shh expression, several breast cell lines, MCF-7, Bcap37, MDA-MB-436 and MDA-MB-231, were used in our experiments. As shown in Figure2a, MCF-7 cells expressed Shh at a relatively higher level, Bcap37 and MDA-MB-231 cells displayed a moderate expression, but MDA-MB-436 cells expressed Shh at a lower level. Interestingly, a similar pattern was shown in NF-κB nuclear expression, suggesting a positive correlation between NF-κB and Shh. As the transcription factor, NF-κB regulates target gene expression in mRNA level. Thus, we also detected the expression of Shh in mRNA level using quantitative RT-PCR. Consistent with protein expression data, Shh was expressed at a higher level in MCF-7 cells, but not in Bcap37, MDA-MB-436 and MDA-MB-231 cells (see Fig.2b), demonstrating that the crucial regulation of Shh expression is presented in the transcription level.

Bottom Line: Comprehensive understanding of the regulation mechanism of Shh in cancer cells is necessary to find an effective approach to selectively block its tumorigenic function.Moreover, in vitro data demonstrated that both NF-κB activation and hypomethylation in promoter region were positively associated with the overexpression of Shh.Furthermore, the biological function data indicated that overexpressed Shh enhanced the self-renewal capacity and migration ability of breast cancer cells, which could be augmented by promoter demethylation and NF-κB activation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, College of Life Science and Biopharmaceutical of Shenyang Pharmaceutical University, Shenyang, China.

Show MeSH
Related in: MedlinePlus