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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 effects of Sonic hedgehog (Shh) overexpression, TNF-α, 5-Aza, cyclopamine or combination on colony formation and migration of breast cancer cells. (a) MM231 colony formation was measured after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 72 h. (b) Bcap37 migration was measured by wound-healing migration assay after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 24 h. All error bars are SEM. *P < 0.05 compare with control group; #P < 0.05 compare with combination group.
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fig04: The effects of Sonic hedgehog (Shh) overexpression, TNF-α, 5-Aza, cyclopamine or combination on colony formation and migration of breast cancer cells. (a) MM231 colony formation was measured after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 72 h. (b) Bcap37 migration was measured by wound-healing migration assay after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 24 h. All error bars are SEM. *P < 0.05 compare with control group; #P < 0.05 compare with combination group.

Mentions: It is well known that the activation of Shh signaling would result in a malignant potential of cancer cells.3,4 Therefore, we next investigated whether hypomethylation, NF-κB and Shh cooperate to drive the malignant potential of breast cancer cells. As shown in Figure4a, the overexpression of Shh led to an increase in the colony formation rate of MDA-MB-231 cells. Similarly, there was a significant increase in the number of colony-forming cells after treatment with TNF-α or 5-Aza. In addition, triple combination resulted in a significant increase in the colony formation rate, indicating an enhanced self-renewal capacity of these breast cancer cells. Interestingly, pretreatment with cyclopamine, a specific inhibitor of Shh signaling, led to an obvious decrease in the number of colony-forming cells, suggesting that the induction of Shh expression by 5-Aza and NF-κB is critical in the self-renewal of breast cancer cells. Moreover, the migration ability of Bcap37 cells was also measured. In agreement with the data obtained from the colony formation assay, the migration cells were increased after being treated with overexpressed Shh, TNF-α or 5-Aza (see Fig.4b). Meanwhile, the number of invasion cells was significantly increased after the combination, but obviously reversed by cyclopamine (see Fig.4b). Furthermore, we also assessed the effect of Shh signaling on cell proliferation. As shown in Figure S1, overexpression of Shh resulted in an increase in the cell viability of MDA-MB-231 cells, whereas pretreatment with cyclopamine reversed Shh-mediated cell proliferation. In contrast to colony formation and migration results, a slight reduction of cell viability was shown after treatment with TNF-α or 5-Aza. Taken together, our data showed that hypomethylation, NF-κB and Shh cooperate to drive self-renewal capacity and migration ability of breast cancer cells.


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 effects of Sonic hedgehog (Shh) overexpression, TNF-α, 5-Aza, cyclopamine or combination on colony formation and migration of breast cancer cells. (a) MM231 colony formation was measured after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 72 h. (b) Bcap37 migration was measured by wound-healing migration assay after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 24 h. All error bars are SEM. *P < 0.05 compare with control group; #P < 0.05 compare with combination group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig04: The effects of Sonic hedgehog (Shh) overexpression, TNF-α, 5-Aza, cyclopamine or combination on colony formation and migration of breast cancer cells. (a) MM231 colony formation was measured after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 72 h. (b) Bcap37 migration was measured by wound-healing migration assay after different agents (5-Aza, pCMV-Shh, TNF-α and cyclopamine) were treated for 24 h. All error bars are SEM. *P < 0.05 compare with control group; #P < 0.05 compare with combination group.
Mentions: It is well known that the activation of Shh signaling would result in a malignant potential of cancer cells.3,4 Therefore, we next investigated whether hypomethylation, NF-κB and Shh cooperate to drive the malignant potential of breast cancer cells. As shown in Figure4a, the overexpression of Shh led to an increase in the colony formation rate of MDA-MB-231 cells. Similarly, there was a significant increase in the number of colony-forming cells after treatment with TNF-α or 5-Aza. In addition, triple combination resulted in a significant increase in the colony formation rate, indicating an enhanced self-renewal capacity of these breast cancer cells. Interestingly, pretreatment with cyclopamine, a specific inhibitor of Shh signaling, led to an obvious decrease in the number of colony-forming cells, suggesting that the induction of Shh expression by 5-Aza and NF-κB is critical in the self-renewal of breast cancer cells. Moreover, the migration ability of Bcap37 cells was also measured. In agreement with the data obtained from the colony formation assay, the migration cells were increased after being treated with overexpressed Shh, TNF-α or 5-Aza (see Fig.4b). Meanwhile, the number of invasion cells was significantly increased after the combination, but obviously reversed by cyclopamine (see Fig.4b). Furthermore, we also assessed the effect of Shh signaling on cell proliferation. As shown in Figure S1, overexpression of Shh resulted in an increase in the cell viability of MDA-MB-231 cells, whereas pretreatment with cyclopamine reversed Shh-mediated cell proliferation. In contrast to colony formation and migration results, a slight reduction of cell viability was shown after treatment with TNF-α or 5-Aza. Taken together, our data showed that hypomethylation, NF-κB and Shh cooperate to drive self-renewal capacity and migration ability of breast cancer cells.

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