Person:
Andrés Gamazo, Paloma Jimena De

First Name

Paloma Jimena De

Last Name

Andrés Gamazo

URI

https://hdl.handle.net/20.500.14352/80475

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Veterinaria

Department

Medicina y Cirugía Animal

Area

Medicina y Cirugía Animal

Identifiers

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Now showing 1 - 8 of 8

Establishment and Characterization of a New Cell Line of Canine Inflammatory Mammary Cancer: IPC-366
(PLoS ONE, 2015) Cáceres Ramos, Sara Cristina; Peña Fernández, Laura Luisa; Andrés Gamazo, Paloma Jimena De; Illera Del Portal, Josefina María; Lopez, Mirtha S.; Woodward, Wendy A.; Reuben, James M.; Illera Del Portal, Juan Carlos
Canine inflammatory mammary cancer (IMC) shares epidemiologic, histopathological and clinical characteristics with the disease in humans and has been proposed as a natural model for human inflammatory breast cancer (IBC). The aim of this study was to characterize a new cell line from IMC (IPC-366) for the comparative study of both IMC and IBC. Tumors cells from a female dog with clinical IMC were collected. The cells were grown under adherent conditions. The growth, cytological, ultrastructural and immunohistochemical (IHC) characteristics of IPC-366 were evaluated. Ten female Balb/SCID mice were inoculated with IPC-366 cells to assess their tumorigenicity and metastatic potential. Chromosome aberration test and Karyotype revealed the presence of structural aberration, numerical and neutral rearrangements, demonstrating a chromosomal instability. Microscopic examination of tumor revealed an epithelial morphology with marked anysocytosis. Cytological and histological examination of smears and ultrathin sections by electron microscopy revealed that IPC-366 is formed by highly malignant large round or polygonal cells characterized by marked atypia and prominent nucleoli and frequent multinucleated cells. Some cells had cytoplasmic empty spaces covered by cytoplasmic membrane resembling capillary endothelial cells, a phenomenon that has been related to s vasculogenic mimicry. IHC characterization of IPC-366 was basal-like: epithelial cells (AE1/AE3+, CK14+, vimentin+, actin-, p63-, ER-, PR-, HER-2, E-cadherin, overexpressed COX-2 and high Ki-67 proliferation index (87.15 %). At 2 weeks after inoculating the IPC-366 cells, a tumor mass was found in 100 % of mice. At 4 weeks metastases in lung and lymph nodes were found. Xenograph tumors maintained the original IHC characteristics of the female dog tumor. In summary, the cell line IPC-366 is a fast growing malignant triple negative cell line model of inflammatory mammary carcinoma that can be used for the comparative study of both IMC and IBC.
Long‐Term Exposure to Isoflavones Alters the Hormonal Steroid Homeostasis‐Impairing Reproductive Function in Adult Male Wistar Rats
(Nutrients, 2023) Cáceres Ramos, Sara Cristina; Crespo, Belen; Alonso Diez, Angela; Andrés Gamazo, Paloma Jimena De; Millán Pastor, María Pilar; Silván Granado, Gema; Illera Del Portal, Josefina María; Illera Del Portal, Juan Carlos
The consumption of isoflavones is gaining popularity worldwide due to their beneficial effects on health. However, isoflavones are considered to be endocrine disruptors and cause deleterious effects on hormone‐sensitive organs, especially in males. Therefore, this study aimed to deter‐ mine if a continuous and prolonged exposure to isoflavones in adult males altered the endocrine axis effect of testicular function. For this purpose, seventy‐five adult male rats were administered with low and high mixtures of isoflavones (genistein and daidzein) for 5 months. The determination of steroid hormones (progesterone, androstenedione, dehydroepiandrosterone, testosterone, dihydrotestosterone, 17β‐estradiol, and estrone sulphate) was carried out in serum and testicular homogenate samples. Sperm quality parameters and testicular histology were also determined. The results revealed that low and high doses of isoflavones promote a hormonal imbalance in androgen and estrogen production, resulting in a decrease in circulating and testicular androgen levels and an increase in estrogen levels. These results are associated with a reduction in the sperm quality parameters and a reduction in the testicular weight, both in the diameter of the seminiferous tubules and the height of the germinal epithelium. Altogether, these results suggest that a continuous ex‐ posure to isoflavones in adult male rats causes a hormonal imbalance in the testes that disrupts the endocrine axis, causing defects in testicular function.
Canine cell line, IPC‐366, as a good model for the study of inflammatory breast cancer
(Veterinary and Comparative Oncology, 2016) Cáceres Ramos, Sara Cristina; Peña Fernández, Laura Luisa; Lacerda, Lara; Illera Del Portal, Josefina María; Andrés Gamazo, Paloma Jimena De; Larson, Richard; Gao, Hui; Debeb, Bisrat; Woodward, Wendy; Reuben, James; Illera Del Portal, Juan Carlos
Inflammatory breast cancer (IBC) is an aggressive type of cancer with poor survival in women. Inflammatory mammary cancer (IMC) in dogs is very similar to human IBC and it has been proposed as a good surrogate model for study the human disease. The aim was to determine if IPC-366 shared characteristics with the IBC cell line SUM149. The comparison was conducted in terms of ability to grow (adherent and nonadherent conditions), stem cell markers expression using flow cytometry, protein production using western blot and tumorigenic capacity. Our results revealed that both are capable of forming long-term mammospheres with a grape-like morphology. Adherent and nonadherent cultures exhibited fast growth in vivo. Stem cell markers expressions showed that IPC-366 and SUM149 in adherent and nonadherent conditions has mesenchymal-like characteristics, E-cadherin and N-cadherin, was higher in adherent than in nonadherent cultures. Therefore, this study determines that both cell lines are similar and IPC-366 is a good model for the human and canine disease.
Predicting Sex in White Rhinoceroses: A Statistical Model for Conservation Management
(Animals, 2023) Martínez, Leticia; Andrés Gamazo, Paloma Jimena De; Caperos, José Manuel; Silván Granado, Gema; Fernández-Morán, Jesús; Casares, Miguel; Crespo, Belén; Vélez Serrano, Daniel; Sanz San Miguel, Luis; Cáceres Ramos, Sara Cristina; Illera Del Portal, Juan Carlos
Ensuring the effective management of every rhinoceros population is crucial for securing a future for the species, especially considering the escalating global threat of poaching and the challenges faced in captive breeding programs for this endangered species. Steroid hormones play pivotal roles in regulating diverse biological processes, making fecal hormonal determinations a valuable non-invasive tool for monitoring adrenal and gonadal endocrinologies and assessing reproductive status, particularly in endangered species. The purpose of this study was to develop a statistical model for predicting the sex of white rhinoceroses using hormonal determinations obtained from a single fecal sample. To achieve this, 562 fecal samples from 15 individuals of the Ceratotherium simum species were collected, and enzyme immunoassays were conducted to determine the concentrations of fecal cortisol, progesterone, estrone, and testosterone metabolites. The biological validation of the method provided an impressive accuracy rate of nearly 80% in predicting the sex of hypothetically unknown white rhinoceroses. Implementing this statistical model for sex identification in white rhinoceroses would yield significant benefits, including a better understanding of the structure and dynamics of wild populations. Additionally, it would enhance conservation management efforts aimed at protecting this endangered species. By utilizing this innovative approach, we can contribute to the preservation and long-term survival of white rhinoceros populations.
In vitro and in vivo effect of flutamide on steroid hormone secretion in canine and human inflammatory breast cancer cell lines
(Veterinary and Comparative Oncology, 2017) Cáceres Ramos, Sara Cristina; Monsalve, Beatriz; Peña Fernández, Laura Luisa; Andrés Gamazo, Paloma Jimena De; Alonso‐Diez, Ángela; Illera Del Portal, Josefina María; Woodward, Wendy; Reuben, James; Silván Granado, Gema; Illera Del Portal, Juan Carlos
The aim was to study the effects of flutamide on cell proliferation, in vivo tumour growth andsteroid production in canine and human IBC cell lines. IPC-366 and SUM149 cell cultures wereexposed to flutamide concentrations for 72 hours. Additionally, IPC-366 and SUM149 xeno-transplanted mice were treated subcutaneously with flutamide 3 times a week for 2 weeks.Steroid hormones determination in culture media, serum and tumour homogenates (pregneno-lone, progesterone, androstenedione, testosterone, dihydrotestosterone, 17β-oestradiol andoestrone sulphate) were assayed by EIA. in vitro cell proliferation percentages showed adecrease in all flutamide dosages in IPC-366 and SUM149. in vivo flutamide reduced tumoursize by 55% to 65%, and metastasis rates decreased. In treated groups, androgen levels in cul-ture media, serum and tumour homogenates were increased as oestrogen levels decreased. These results suggest that flutamide treatment inhibits cell proliferation and promotes tumourreduction by increasing androgen levels and also support future therapy approaches
Non-Invasive Determination of Annual Fecal Cortisol, Androstenedione, and Testosterone Variations in a Herd of Male Asian Elephants (Elephas maximus) and Their Relation to Some Climatic Variables
(Animals, 2021) Andrés Gamazo, Paloma Jimena De; Cáceres Ramos, Sara Cristina; Belén Crespo; Silván Granado, Gema; Illera Del Portal, Juan Carlos
The measurement of stress and reproductive hormones in wild animal species by non-invasive methods is of special interest. To assess whether the adrenal and gonadal hormones show annual variations in male Asian elephants (Elephas maximus) and to evaluate whether there is any influence of climatic variables on hormonal secretion, fecal samples were taken from a herd of 7 Asian elephants over a 14-month period to subsequently determine the concentrations of testosterone (T), androstenedione (A4), and cortisol (C) by a validated immunoassay technique. Data referring to three climatic variables in the place and period of study were collected, namely monthly mean values of temperature, humidity and rainfall. Levels of T and A4 showed two major increases in July (T: 1088.35 ± 131.04 ng/g; A4: 480.40 ± 50.86 ng/g) and October (T: 825.09 ± 31.60 ng/g; A4: 319.96 ± 32.69 ng/g) (p < 0.05). Our results show a secretion of fecal androgens dependent on temperature (T and A4), and humidity (T). Male musth was detected during the initial increases of T and A4 levels. The highest concentrations of C were observed in September (156.67 ± 60.89 ng/g) (p < 0.05), probably due to the stressful fights that occurred during the musth period. The observed results of the fecal levels of T, A4 and C were similar to those obtained by invasive methods. In conclusion, fecal secretion of the three hormones in these captive male Asian elephants showed variations related in some cases to different weather factors.
Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines
(Veterinary Sciences, 2021) Cáceres Ramos, Sara Cristina; Alonso-Diez, Angela; Crespo, Belén; Peña Fernández, Laura Luisa; Illera Del Portal, Josefina María; Silván Granado, Gema; Andrés Gamazo, Paloma Jimena De; Illera Del Portal, Juan Carlos
Xenografts can grow in immunosuppressed hosts, such as SCID mice, and tumor material can be injected into hosts either ectopically or orthotopically. Choosing the correct model to use is a crucial step in animal research. The aim of this study was to report the differences between ectopic and orthotopic xenografts in tumor progression, metastasis capacity, histological features, and steroid hormone profiles in xenografts from the cIMC (canine inflammatory mammary cancer) cell line IPC-366 and hIBC (human inflammatory breast cancer) cell line SUM149. To achieve this purpose, 40 female mice 6–8 weeks old were inoculated with IPC-366 and SUM149 cells subcutaneously (ectopic models) or into mammary fat pad (orthotopic models). Mice were monitored for tumor progression and appearance of metastases, and generated tumors were analyzed in terms of histological examination and steroid hormone production. The results revealed differences in tumor appearance and percentage of metastasis between ectopic and orthotopic models, which were higher in the ectopic xenografts from both cell lines. However, both models had similar characteristics of tumor progression, histological features, and steroid hormone secretion profiles. We show that the ectopic model can be validated as a good and useful model of tumor development in addition to, not contrary to, the orthotopic model in breast cancer research.
Blocking Estrogen Synthesis Leads to Different Hormonal Responses in Canine and Human Triple Negative Inflammatory Breast Cancer
(Cancers, 2021) Cáceres Ramos, Sara Cristina; Beatriz Monsalve; Angela Alonso-Diez; Belén Crespo; Illera Del Portal, Josefina María; Andrés Gamazo, Paloma Jimena De; Silván Granado, Gema; Illera Del Portal, Juan Carlos
Blocking estrogen synthesis by inhibitors of estrogen synthesis is a widely used therapy against estrogen receptor-positive tumors. However, these therapies are less effective in negative expression tumors. Therefore, this study determined the effectiveness of anti-aromatase and anti-sulfatase therapies in canine and human inflammatory breast cancer. Cell cultures and xenografts from IPC-366 and SUM149 were treated with different doses of letrozole (anti-aromatase) and STX-64 (anti-sulfatase), in order to observe their effectiveness in terms of cell proliferation, tumor progression, and the appearance of metastases and hormonal profiles. The results revealed that both treatments are effective in vitro since they reduce cell proliferation and decrease the secreted estrogen levels. In xenograft mice, while treatment with letrozole reduces tumor progression by 30-40%, STX-64 increases tumor progression by 20%. The hormonal results obtained determined that STX-64 produced an increase in circulating and intratumoral levels of estradiol, which led to an increase in tumor progression. However, letrozole was able to block estrogen synthesis by decreasing the levels of circulating and intratumoral estrogen and thus slowing down tumor progression. In conclusion, letrozole can be an effective treatment for canine and human inflammatory breast cancer. The knowledge of the hormonal profile of breast tumors reflects useful information on the effectiveness of different endocrine treatments.