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Estrogen Receptor β Agonists Differentially Affect the Growth of Human Melanoma Cell Lines.

Marzagalli M, Casati L, Moretti RM, Montagnani Marelli M, Limonta P - PLoS ONE (2015)

Bottom Line: Moreover, the antiproliferative activity of ERβ agonists was associated with an altered expression of G1-S transition-related proteins.In these cells, global DNA was found to be hypomethylated when compared to normal melanocytes; this DNA hypomethylation status was reverted by ERβ activation.ERβ agonists also decreased the proliferation of WM115 (BRAF V600D-mutant) cells, while they failed to reduce the growth of A375 and WM1552 (BRAF V600E-mutant) cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milano, Italy.

ABSTRACT

Background: Cutaneous melanoma is an aggressive malignancy; its incidence is increasing worldwide and its prognosis remains poor. Clinical observations indicate that estrogen receptor β (ERβ) is expressed in melanoma tissues and its expression decreases with tumor progression, suggesting its tumor suppressive function. These experiments were performed to investigate the effects of ERβ activation on melanoma cell growth.

Methods and results: Protein expression was analyzed by Western blot and immunofluorescence assays. Cell proliferation was assessed by counting the cells by hemocytometer. ERβ transcriptional activity was evaluated by gene reporter assay. Global DNA methylation was analyzed by restriction enzyme assay and ERβ isoforms were identified by qRT-PCR. We demonstrated that ERβ is expressed in a panel of human melanoma cell lines (BLM, WM115, A375, WM1552). In BLM (NRAS-mutant) cells, ERβ agonists significantly and specifically inhibited cell proliferation. ERβ activation triggered its cytoplasmic-to-nuclear translocation and transcriptional activity. Moreover, the antiproliferative activity of ERβ agonists was associated with an altered expression of G1-S transition-related proteins. In these cells, global DNA was found to be hypomethylated when compared to normal melanocytes; this DNA hypomethylation status was reverted by ERβ activation. ERβ agonists also decreased the proliferation of WM115 (BRAF V600D-mutant) cells, while they failed to reduce the growth of A375 and WM1552 (BRAF V600E-mutant) cells. Finally, we could observe that ERβ isoforms are expressed at different levels in the various cell lines. Specific oncogenic mutations or differential expression of receptor isoforms might be responsible for the different responses of cell lines to ERβ agonists.

Conclusions: Our results demonstrate that ERβ is expressed in melanoma cell lines and that ERβ agonists differentially regulate the proliferation of these cells. These data confirm the notion that melanoma is a heterogeneous tumor and that genetic profiling is mandatory for the development of effective personalized therapeutic approaches for melanoma patients.

No MeSH data available.


Related in: MedlinePlus

ERβ isoforms (1, 2, and 5) are differentially expressed in melanoma cell lines.The relative expression of ERβ1, 2, and 5 isoforms was evaluated by quantitative RT-PCR, utilizing specific sets of primers. BLM and WM115 cells showed a similar expression of ERβ1 and 5, while expressing higher levels of ERβ2. On the other hand, a high expression of both ERβ2 and 5 isoforms (when compared to ERβ 1) was observed in A375 cells. One representative of three different experiments, which gave similar results, is shown.
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pone.0134396.g007: ERβ isoforms (1, 2, and 5) are differentially expressed in melanoma cell lines.The relative expression of ERβ1, 2, and 5 isoforms was evaluated by quantitative RT-PCR, utilizing specific sets of primers. BLM and WM115 cells showed a similar expression of ERβ1 and 5, while expressing higher levels of ERβ2. On the other hand, a high expression of both ERβ2 and 5 isoforms (when compared to ERβ 1) was observed in A375 cells. One representative of three different experiments, which gave similar results, is shown.

Mentions: Fig 7 shows that the pattern of expression of the ERβ isoforms is similar in BLM and in WM115 cells, with ERβ1and ERβ5 being expressed at similar levels and ERβ2 showing a higher level of expression. On the other hand, in A375 cells, both ERβ2 and 5 are expressed at higher levels than the ERβ1 isoform.


Estrogen Receptor β Agonists Differentially Affect the Growth of Human Melanoma Cell Lines.

Marzagalli M, Casati L, Moretti RM, Montagnani Marelli M, Limonta P - PLoS ONE (2015)

ERβ isoforms (1, 2, and 5) are differentially expressed in melanoma cell lines.The relative expression of ERβ1, 2, and 5 isoforms was evaluated by quantitative RT-PCR, utilizing specific sets of primers. BLM and WM115 cells showed a similar expression of ERβ1 and 5, while expressing higher levels of ERβ2. On the other hand, a high expression of both ERβ2 and 5 isoforms (when compared to ERβ 1) was observed in A375 cells. One representative of three different experiments, which gave similar results, is shown.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4520550&req=5

pone.0134396.g007: ERβ isoforms (1, 2, and 5) are differentially expressed in melanoma cell lines.The relative expression of ERβ1, 2, and 5 isoforms was evaluated by quantitative RT-PCR, utilizing specific sets of primers. BLM and WM115 cells showed a similar expression of ERβ1 and 5, while expressing higher levels of ERβ2. On the other hand, a high expression of both ERβ2 and 5 isoforms (when compared to ERβ 1) was observed in A375 cells. One representative of three different experiments, which gave similar results, is shown.
Mentions: Fig 7 shows that the pattern of expression of the ERβ isoforms is similar in BLM and in WM115 cells, with ERβ1and ERβ5 being expressed at similar levels and ERβ2 showing a higher level of expression. On the other hand, in A375 cells, both ERβ2 and 5 are expressed at higher levels than the ERβ1 isoform.

Bottom Line: Moreover, the antiproliferative activity of ERβ agonists was associated with an altered expression of G1-S transition-related proteins.In these cells, global DNA was found to be hypomethylated when compared to normal melanocytes; this DNA hypomethylation status was reverted by ERβ activation.ERβ agonists also decreased the proliferation of WM115 (BRAF V600D-mutant) cells, while they failed to reduce the growth of A375 and WM1552 (BRAF V600E-mutant) cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milano, Italy.

ABSTRACT

Background: Cutaneous melanoma is an aggressive malignancy; its incidence is increasing worldwide and its prognosis remains poor. Clinical observations indicate that estrogen receptor β (ERβ) is expressed in melanoma tissues and its expression decreases with tumor progression, suggesting its tumor suppressive function. These experiments were performed to investigate the effects of ERβ activation on melanoma cell growth.

Methods and results: Protein expression was analyzed by Western blot and immunofluorescence assays. Cell proliferation was assessed by counting the cells by hemocytometer. ERβ transcriptional activity was evaluated by gene reporter assay. Global DNA methylation was analyzed by restriction enzyme assay and ERβ isoforms were identified by qRT-PCR. We demonstrated that ERβ is expressed in a panel of human melanoma cell lines (BLM, WM115, A375, WM1552). In BLM (NRAS-mutant) cells, ERβ agonists significantly and specifically inhibited cell proliferation. ERβ activation triggered its cytoplasmic-to-nuclear translocation and transcriptional activity. Moreover, the antiproliferative activity of ERβ agonists was associated with an altered expression of G1-S transition-related proteins. In these cells, global DNA was found to be hypomethylated when compared to normal melanocytes; this DNA hypomethylation status was reverted by ERβ activation. ERβ agonists also decreased the proliferation of WM115 (BRAF V600D-mutant) cells, while they failed to reduce the growth of A375 and WM1552 (BRAF V600E-mutant) cells. Finally, we could observe that ERβ isoforms are expressed at different levels in the various cell lines. Specific oncogenic mutations or differential expression of receptor isoforms might be responsible for the different responses of cell lines to ERβ agonists.

Conclusions: Our results demonstrate that ERβ is expressed in melanoma cell lines and that ERβ agonists differentially regulate the proliferation of these cells. These data confirm the notion that melanoma is a heterogeneous tumor and that genetic profiling is mandatory for the development of effective personalized therapeutic approaches for melanoma patients.

No MeSH data available.


Related in: MedlinePlus