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Melatonin Attenuates Her-2, p38 MAPK, p-AKT, and mTOR Levels in Ovarian Carcinoma of Ethanol-Preferring Rats.

Ferreira GM, Martinez M, Camargo IC, Domeniconi RF, Martinez FE, Chuffa LG - J Cancer (2014)

Bottom Line: While Mel therapy was unable to reduce Her-4 and phosphoinositide 3-kinase (PI3K) levels, it was able to suppress the OC-related increase in the levels of the Her-2, p38 mitogen-activated protein kinases (p38 MAPK), protein kinase B (phospho-AKT), and mammalian target of rapamycin (mTOR).In addition, Mel significantly attenuated the expression of Her-2, p38 MAPK, and p-AKT, which are involved in OC signaling during ethanol intake.Collectively, our results suggest that Mel attenuates the Her-2-signaling pathway in OC of ethanol-preferring rats, providing an effective contribution for further development of adjuvant therapies.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Anatomy, Biosciences Institute, UNESP - Univ. Estadual Paulista, Botucatu-SP, Brazil, 18618-970.

ABSTRACT
Epidermal growth factor receptors 2 (Her-2) and 4 (Her-4) are closely associated with ovarian cancer (OC) progression and metastasis, and a more complete understanding of these signaling pathways allow the development of new therapeutic strategies. Melatonin (Mel) is recognized as having several anticancer properties and has been reported to modulate Her-2 system in aggressive tumors. Here, we investigated OC and the role of Mel therapy on the Her-2- and Her-4-signaling pathway related to downstream molecules in an ethanol-preferring rat model. To induce OC, the left ovary was injected directly with a single dose of 100 µg 7,12-dimethylbenz(a)anthracene (DMBA) dissolved in 10 µL of sesame oil under the bursa. Right ovaries were used as sham-surgery controls. After developing OC, half of the animals received i.p. injections of Mel (200 µg/100 g b.w./day) for 60 days. While Mel therapy was unable to reduce Her-4 and phosphoinositide 3-kinase (PI3K) levels, it was able to suppress the OC-related increase in the levels of the Her-2, p38 mitogen-activated protein kinases (p38 MAPK), protein kinase B (phospho-AKT), and mammalian target of rapamycin (mTOR). In addition, Mel significantly attenuated the expression of Her-2, p38 MAPK, and p-AKT, which are involved in OC signaling during ethanol intake. Collectively, our results suggest that Mel attenuates the Her-2-signaling pathway in OC of ethanol-preferring rats, providing an effective contribution for further development of adjuvant therapies.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical/fluorescence localization and Western blot analysis of Her-2 and Her-4 in serous papillary OC. The immunoreaction of Her-2 was intense in the surface epithelium of the OC (A) and OC+EtOH (C) groups (arrows), but not in the stroma. Absence of reaction to Her-2 was observed in the OC+Mel (B) group (arrow), while a weak Her-2 reaction appeared in the epithelium of the OC+EtOH+Mel (D) group (arrow). A moderate Her-4 reaction was similar in the epithelium of the OC (E), OC+Mel (F), OC+EtOH (G), and OC+EtOH+Mel (H) groups (arrow). Bar = 20 µm. Negative controls were used. (I) Representative Her-2 and Her-4 profiles of extracts (70 µg protein) pooled from 10 samples/group (upper panel). (J-K) Extracts obtained from individual animals were used for densitometric analysis of the Her-2 and Her-4 levels following normalization to the β-actin. All results are expressed as the mean ± SD (n = 10). a P < 0.05 versus OC; b P < 0.05 versus OC+EtOH. (L-O) Merged images of the immunofluorescence of Her-2 and DAPI nuclear staining in OC (L), OC+Mel (M), OC+EtOH (N) and OC+EtOH+Mel (O); details of the cell receptor staining (Alexa fluor® 488, Bar = 10 µm).
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Figure 3: Immunohistochemical/fluorescence localization and Western blot analysis of Her-2 and Her-4 in serous papillary OC. The immunoreaction of Her-2 was intense in the surface epithelium of the OC (A) and OC+EtOH (C) groups (arrows), but not in the stroma. Absence of reaction to Her-2 was observed in the OC+Mel (B) group (arrow), while a weak Her-2 reaction appeared in the epithelium of the OC+EtOH+Mel (D) group (arrow). A moderate Her-4 reaction was similar in the epithelium of the OC (E), OC+Mel (F), OC+EtOH (G), and OC+EtOH+Mel (H) groups (arrow). Bar = 20 µm. Negative controls were used. (I) Representative Her-2 and Her-4 profiles of extracts (70 µg protein) pooled from 10 samples/group (upper panel). (J-K) Extracts obtained from individual animals were used for densitometric analysis of the Her-2 and Her-4 levels following normalization to the β-actin. All results are expressed as the mean ± SD (n = 10). a P < 0.05 versus OC; b P < 0.05 versus OC+EtOH. (L-O) Merged images of the immunofluorescence of Her-2 and DAPI nuclear staining in OC (L), OC+Mel (M), OC+EtOH (N) and OC+EtOH+Mel (O); details of the cell receptor staining (Alexa fluor® 488, Bar = 10 µm).

Mentions: To validate the treatment, plasma Mel levels were measured at 30- and 60-day treatment, and efficiently, OC+Mel and OC+EtOH+Mel groups showed increased levels of circulating Mel (Fig. 2). In response to Mel therapy, the expression and immunostaining of Her-2 was decreased in the OC+Mel group (1.59-fold reduced vs. OC group) and in the OC+EtOH+Mel group (1.33-fold reduced vs. OC+EtOH group) in serous papillary OC (Fig. 3 A-D, I, J). In contrast, the expression of Her-4 was unchanged along the treatments (Fig. 3 E-H, I, K). Immunofluorescence assays detected the level and localization of Her-2 in OC cells. Mel therapy resulted in downregulation of Her-2 expression (Fig. 3 L, M; fluorescence level reduced from 88% ± 9.2 (OC) to 46% ± 11.6 (OC+Mel)). Interestingly, EtOH intake significantly increased Her-2 levels in OC, and Mel therapy drastically decreased these levels (Fig. 3 N, O; fluorescence level reduced from 113% ± 13.2 (OC+EtOH) to 63% ± 10.1 (OC+EtOH+Mel)).


Melatonin Attenuates Her-2, p38 MAPK, p-AKT, and mTOR Levels in Ovarian Carcinoma of Ethanol-Preferring Rats.

Ferreira GM, Martinez M, Camargo IC, Domeniconi RF, Martinez FE, Chuffa LG - J Cancer (2014)

Immunohistochemical/fluorescence localization and Western blot analysis of Her-2 and Her-4 in serous papillary OC. The immunoreaction of Her-2 was intense in the surface epithelium of the OC (A) and OC+EtOH (C) groups (arrows), but not in the stroma. Absence of reaction to Her-2 was observed in the OC+Mel (B) group (arrow), while a weak Her-2 reaction appeared in the epithelium of the OC+EtOH+Mel (D) group (arrow). A moderate Her-4 reaction was similar in the epithelium of the OC (E), OC+Mel (F), OC+EtOH (G), and OC+EtOH+Mel (H) groups (arrow). Bar = 20 µm. Negative controls were used. (I) Representative Her-2 and Her-4 profiles of extracts (70 µg protein) pooled from 10 samples/group (upper panel). (J-K) Extracts obtained from individual animals were used for densitometric analysis of the Her-2 and Her-4 levels following normalization to the β-actin. All results are expressed as the mean ± SD (n = 10). a P < 0.05 versus OC; b P < 0.05 versus OC+EtOH. (L-O) Merged images of the immunofluorescence of Her-2 and DAPI nuclear staining in OC (L), OC+Mel (M), OC+EtOH (N) and OC+EtOH+Mel (O); details of the cell receptor staining (Alexa fluor® 488, Bar = 10 µm).
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Figure 3: Immunohistochemical/fluorescence localization and Western blot analysis of Her-2 and Her-4 in serous papillary OC. The immunoreaction of Her-2 was intense in the surface epithelium of the OC (A) and OC+EtOH (C) groups (arrows), but not in the stroma. Absence of reaction to Her-2 was observed in the OC+Mel (B) group (arrow), while a weak Her-2 reaction appeared in the epithelium of the OC+EtOH+Mel (D) group (arrow). A moderate Her-4 reaction was similar in the epithelium of the OC (E), OC+Mel (F), OC+EtOH (G), and OC+EtOH+Mel (H) groups (arrow). Bar = 20 µm. Negative controls were used. (I) Representative Her-2 and Her-4 profiles of extracts (70 µg protein) pooled from 10 samples/group (upper panel). (J-K) Extracts obtained from individual animals were used for densitometric analysis of the Her-2 and Her-4 levels following normalization to the β-actin. All results are expressed as the mean ± SD (n = 10). a P < 0.05 versus OC; b P < 0.05 versus OC+EtOH. (L-O) Merged images of the immunofluorescence of Her-2 and DAPI nuclear staining in OC (L), OC+Mel (M), OC+EtOH (N) and OC+EtOH+Mel (O); details of the cell receptor staining (Alexa fluor® 488, Bar = 10 µm).
Mentions: To validate the treatment, plasma Mel levels were measured at 30- and 60-day treatment, and efficiently, OC+Mel and OC+EtOH+Mel groups showed increased levels of circulating Mel (Fig. 2). In response to Mel therapy, the expression and immunostaining of Her-2 was decreased in the OC+Mel group (1.59-fold reduced vs. OC group) and in the OC+EtOH+Mel group (1.33-fold reduced vs. OC+EtOH group) in serous papillary OC (Fig. 3 A-D, I, J). In contrast, the expression of Her-4 was unchanged along the treatments (Fig. 3 E-H, I, K). Immunofluorescence assays detected the level and localization of Her-2 in OC cells. Mel therapy resulted in downregulation of Her-2 expression (Fig. 3 L, M; fluorescence level reduced from 88% ± 9.2 (OC) to 46% ± 11.6 (OC+Mel)). Interestingly, EtOH intake significantly increased Her-2 levels in OC, and Mel therapy drastically decreased these levels (Fig. 3 N, O; fluorescence level reduced from 113% ± 13.2 (OC+EtOH) to 63% ± 10.1 (OC+EtOH+Mel)).

Bottom Line: While Mel therapy was unable to reduce Her-4 and phosphoinositide 3-kinase (PI3K) levels, it was able to suppress the OC-related increase in the levels of the Her-2, p38 mitogen-activated protein kinases (p38 MAPK), protein kinase B (phospho-AKT), and mammalian target of rapamycin (mTOR).In addition, Mel significantly attenuated the expression of Her-2, p38 MAPK, and p-AKT, which are involved in OC signaling during ethanol intake.Collectively, our results suggest that Mel attenuates the Her-2-signaling pathway in OC of ethanol-preferring rats, providing an effective contribution for further development of adjuvant therapies.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Anatomy, Biosciences Institute, UNESP - Univ. Estadual Paulista, Botucatu-SP, Brazil, 18618-970.

ABSTRACT
Epidermal growth factor receptors 2 (Her-2) and 4 (Her-4) are closely associated with ovarian cancer (OC) progression and metastasis, and a more complete understanding of these signaling pathways allow the development of new therapeutic strategies. Melatonin (Mel) is recognized as having several anticancer properties and has been reported to modulate Her-2 system in aggressive tumors. Here, we investigated OC and the role of Mel therapy on the Her-2- and Her-4-signaling pathway related to downstream molecules in an ethanol-preferring rat model. To induce OC, the left ovary was injected directly with a single dose of 100 µg 7,12-dimethylbenz(a)anthracene (DMBA) dissolved in 10 µL of sesame oil under the bursa. Right ovaries were used as sham-surgery controls. After developing OC, half of the animals received i.p. injections of Mel (200 µg/100 g b.w./day) for 60 days. While Mel therapy was unable to reduce Her-4 and phosphoinositide 3-kinase (PI3K) levels, it was able to suppress the OC-related increase in the levels of the Her-2, p38 mitogen-activated protein kinases (p38 MAPK), protein kinase B (phospho-AKT), and mammalian target of rapamycin (mTOR). In addition, Mel significantly attenuated the expression of Her-2, p38 MAPK, and p-AKT, which are involved in OC signaling during ethanol intake. Collectively, our results suggest that Mel attenuates the Her-2-signaling pathway in OC of ethanol-preferring rats, providing an effective contribution for further development of adjuvant therapies.

No MeSH data available.


Related in: MedlinePlus