Pigment epithelium derived factor (PEDF) expression in the male tract of Wistar rats.

Pigment epithelium derived factor (PEDF) expression has been described in many organs as showing neurotrophic, anti-angiogenic, anti-apoptotic, anti-inflammatory, anti-oxidant and pro-cell survival properties. However, references to its activity in the male reproductive system are scarce. We aimed to characterize the expression of PEDF in the male reproductive tract of Wistar rats by using RT-PCR, western blot and immunostaining and also evaluate the effect of flutamide in PEDF expression. We found that PEDF is expressed in the epididymis, prostate and seminal vesicles in Wistar rats, but notably not in the testes. Under the effect of flutamide PEDF expression decreased, recovering by suppressing the antiandrogen. The epididymis is an essential organ in sperm maturation-storages. The role of PEDF in this physiological process has not been fully elucidated yet, but considering that in other systems PEDF has anti-apoptotic, anti-oxidants and pro-cell survival properties, its expression along the epididymis could play a role in the protection of spermatozoa while they are stored.

Molecular markers of DNA damage and repair in cervical cancer patients treated with cisplatin neoadjuvant chemotherapy: an exploratory study.

Neoadjuvant (or induction) chemotherapy can be used for cervical cancer patients with locally advanced disease; this treatment is followed by radical surgery and/or radiation therapy. Cisplatin is considered to be the most active platinum agent drug for this cancer, with a response rate of 20%. In order to understand how the cisplatin treatment affects the stress response, in this work, we performed an exploratory study to analyze a number of stress proteins before and after cisplatin neoadjuvant chemotherapy. The study involved 14 patients; the pre- and post-chemotherapy paired biopsies were examined by hematoxylin and eosin staining and by immunohistochemistry. The proteins evaluated were p53, P16/INK4A, MSH2, nuclear protein transcriptional regulator 1 (NUPR1), and HSPB1 (total: HSPB1/t and phosphorylated: HSPB1/p). These proteins were selected because there is previous evidence of their relationship with drug resistance. The formation of platinum-DNA adducts was also studied. There was a great variation in the expression levels of the mentioned proteins in the pre-chemotherapy biopsies. After chemotherapy, p53 was not significantly affected by cisplatin, as well as P16/INK4A and MSH2 while nuclear NUPR1 content tended to decrease (p = 0.056). Cytoplasmic HSPB1/t expression levels decreased significantly following cisplatin therapy while nuclear HSPB1/t and HSPB1/p tended to increase. Since the most significant changes following chemotherapy appeared in the HSPB1 expression levels, the changes were confirmed by Western blot. The platinum-DNA adducts were observed in HeLa cell in apoptosis; however, in the tumor samples, the platinum-DNA adducts were observed in morphologically healthy tumor cells; these cells displayed nuclear HSPB1/p. Further mechanistic studies should be performed to reveal how HSPB1/p is related with drug resistance. When the correlations of the markers with the response to neoadjuvant chemotherapy were examined, only high pre-chemotherapy levels of cytoplasmic HSPB1/p correlated with a poor clinical and pathological response to neoadjuvant cisplatin chemotherapy (p = 0.056) suggesting that this marker could be useful opening its study in a larger number of cases.

HER2 and ß-catenin protein location: importance in the prognosis of breast cancer patients and their correlation when breast cancer cells suffer stressful situations.

In human breast cancer, ß-catenin localization has been related with disease prognosis. Since HER2-positive patients are an important subgroup, and that in breast cancer cells a direct interaction of ß-catenin/HER2 has been reported, in the present study we have explored whether ß-catenin location is related with the disease survival. The study was performed in a tumor bank from patients (n = 140) that did not receive specific anti-HER2 therapy. The proteins were detected by immunohistochemistry in serial sections, 47 (33.5%) patients were HER2-positive with a long follow-up. HER2-positive patients that displayed ß-catenin at the plasma membrane (completely surrounding the tumour cells) showed a significant better disease-free survival and overall survival than the patients showing the protein on other locations. Then we explored the dynamics of the co-expression of ß-catenin and HER2 in human MCF-7 and SKBR3 cells exposed to different stressful situations. In untreated conditions MCF-7 and SKBR3 cells showed very different ß-catenin localization. In MCF-7 cells, cadmium administration caused a striking change in ß-catenin localization driving it from plasma membrane to cytoplasmic and perinuclear areas and HER2 showed a similar localization patterns. The changes induced by cadmium were compared with heat shock, H2O2 and tamoxifen treatments. In conclusion, this study shows the dynamical associations of HER2 and ß-catenin and their changes in subcellular localizations driven by stressful situations. In addition, we report for the first time the correlation between plasma membrane associated ß-catenin in HER2-positive breast cancer and survival outcome, and the importance of the protein localization in breast cancer samples.

Effects of temozolomide (TMZ) on the expression and interaction of heat shock proteins (HSPs) and DNA repair proteins in human malignant glioma cells.

We previously reported the association of HSPA1A and HSPB1 with high-grade astrocytomas, suggesting that these proteins might be involved in disease outcome and response to treatment. With the aim to better understand the resistance/susceptibility processes associated to temozolomide (TMZ) treatment, the current study was performed in three human malignant glioma cell lines by focusing on several levels: (a) apoptotic index and senescence, (b) DNA damage, and (c) interaction of HSPB1 with players of the DNA damage response. Three human glioma cell lines, Gli36, U87, and DBTRG, were treated with TMZ evaluating cell viability and survival, apoptosis, senescence, and comets (comet assay). The expression of HSPA (HSPA1A and HSPA8), HSPB1, O6-methylguanine-DNA methyltransferase (MGMT), MLH1, and MSH2 was determined by immunocytochemistry, immunofluorescence, and Western blot. Immunoprecipitation was used to analyze protein interaction. The cell lines exhibited differences in viability, apoptosis, and senescence after TMZ administration. We then focused on Gli36 cells (relatively unstudied) which showed very low recovery capacity following TMZ treatment, and this was related to high DNA damage levels; however, the cells maintained their viability. In these cells, MGMT, MSH2, HSPA, and HSPB1 levels increased significantly after TMZ administration. In addition, MSH2 and HSPB1 proteins appeared co-localized by confocal microscopy. This co-localization increased after TMZ treatment, and in immunoprecipitation analysis, MSH2 and HSPB1 appeared interacting. In contrast, HSPB1 did not interact with MGMT. We show in glioma cells the biological effects of TMZ and how this drug affects the expression levels of heat shock proteins (HSPs), MGMT, MSH2, and MLH1. In Gli36 cells, the results suggest that interactions between HSPB1 and MSH2, including co-nuclear localization, may be important in determining cell sensitivity to TMZ.

In MMTV-Her-2/neu transgenic mammary tumors the absence of caveolin-1-/- alters PTEN and NHERF1 but not ß-catenin expression.

In a recent study, we have shown that in mammary tumors from mice lacking the Cav-1 gene, there are alterations in specific heat shock proteins as well as in tumor development. With this in mind, we have now investigated other proteins in the same mammary mouse tumor model (Her-2/neu expressing mammary tumors from Cav-1 wild type and Cav-1 null mice), to further comprehend the complex tumor-stroma mechanisms involved in regulating stress responses during tumor development. In this tumor model the cancer cells always lacked of Cav-1, so the KO influenced the Cav-1 in the stroma. By immunohistochemistry, we have found a striking co-expression of ß-catenin and Her-2/neu in the tumor cells. The absence of Cav-1 in the tumor stroma had no effect on expression or localization of ß-catenin and Her-2/neu. Both proteins appeared co-localized at the cell surface during tumor development and progression. Since Her-2/neu activation induces MTA1, we next evaluated MTA1 in the mouse tumors. Although this protein was found in numerous nuclei, the absence of Cav-1 did not alter its expression level. In contrast, significantly more PTEN protein was noted in the tumors lacking Cav-1 in the stroma, with the protein localized mainly in the nuclei. P-Akt levels were relatively low in tumors from both Cav-1 WT and Cav-1 KO mice. There was also an increase in nuclear NHERF1 expression levels in the tumors arising from Cav-1 KO mice. The data obtained in the MMTV-neu model are consistent with a role for Cav-1 in adjacent breast cancer stromal cells in modulating the expression and localization of important proteins implicated in tumor cell behavior.

Downregulation of Hsp27 (HSPB1) in MCF-7 human breast cancer cells induces upregulation of PTEN.

Hsp27 (HSPB1) is usually overexpressed in breast cancers affecting the disease outcome and the sensitivity of tumors to chemotherapy and radiotherapy. Hsp27 interacts with other proteins such as ß-catenin, histone deacetylase HDAC6, transcription factor STAT2 and procaspase-3. Phosphatase and tensin homologue (PTEN) is a tumor suppressor gene that is deleted in many human tumors. The PI3K/Akt signaling pathway is negatively regulated by PTEN. Hsp27 is described as a key component of the Akt signaling cascade: Akt, BAD, Forkhead transcription factors, Hsp27, mitogen-activated protein kinase kinase-3 and -6. Here, we have examined whether the downregulation of Hsp27 by siHsp27 affects the PTEN levels in the MCF-7 human breast cancer cell line. PTEN was detected with two different antibodies using western blots and immunocytochemistry. p-Akt was also evaluated by western blot. In addition, Hsp27 and PTEN were immunoprecipitated to know whether these proteins interact. Intracellular colocalization studies were carried out by confocal microscopy. A significant reduction in the Hsp27 levels was noted in the siHsp27 transfected cells. These Hsp27 downregulated cells showed a significant increased expression of PTEN. The MW 76 and 55 kDa PTEN forms were upregulated as revealed by two different antibodies. The phosphatase activity of PTEN seems to be active because p-Akt levels were reduced. Hsp27 immunoprecipitation was bringing PTEN and vice versa, these two proteins seem to interact at cytoplasmic level by FRET. Downregulation of Hsp27 stabilized PTEN protein levels. Chaperone-assisted E3 ligase C terminus of Hsc70-interacting protein (CHIP) levels were not significantly influenced by Hsp27 downregulation. In conclusion, we report a novel function of Hsp27 modulating the PTEN levels in human breast cancer cells suggesting an interaction between these two molecules.

Hsp27 (HSPB1): a possible surrogate molecular marker for loss of heterozygosity (LOH) of chromosome 1p in oligodendrogliomas but not in astrocytomas.

In oligodendrogliomas, 1p loss of heterozygosity (LOH) is a predictor of good prognosis and treatment response. In contrast, in uveal melanomas, LOH of chromosome 3 has been linked to poor prognosis and downregulation of Hsp27. In the present study, we have analyzed the expression of heat-shock proteins (Hsps) to characterize subtypes of gliomas and their histopathologic features and to correlate with other molecular markers including LOH of 1p. Biopsies from patients with primary gliomas (n = 65) were analyzed by immunohistochemistry, chromogenic in situ hybridization and fluorescent in situ hybridization and methylation-specific PCR (MSP). Elevated Hsp27 and total Hsp70 expression levels were associated with high-grade astrocytomas (p = 0.0001 and p = 0.01, respectively). In grade III oligodendrogliomas, the Hsp27 levels were significantly higher (p = 0.03). Low O6-methylguanine-DNA methyltransferase (MGMT) expression was associated with grade II astrocytomas. Elevated ß-catenin expression was associated with grade III/IV astrocytomas (p = 0.003); p53 (+) tumors were more frequently found in grade III/IV astrocytomas (p = 0,001). LOH on 1p was associated with oligodendroglial tumours. In addition, a higher Hsp27 expression correlated with LOH of 1p (p = 0.017); this was also tested in two glioma cell lines. MSP was successful in only six samples. No significant correlations were found for the other markers. In conclusion, in oligodendroglial tumors, Hsp27 appeared as a surrogate marker of LOH of 1p which could also help to predict the disease prognosis. In gliomas, p53, Hsp27, Hsp70, MGMT, and ß-catenin correlated with histopathological characteristics, suggesting that these markers could predict the disease outcome and the response to treatments.

Isolation of heat shock protein complexes.

Heat shock proteins (Hsp) are molecular chaperones with the capability to interact with a wide range of other proteins and are thus often found coupled with other heat shock and non-heat shock proteins. This can be an advantage to study specific interactions between a chaperone and other proteins and to generate an antitumoral immune response. In this chapter, we present two protocols to isolate Hsp. One involves column chromatography with hydroxyapatite and the other employs immunoprecipitation with antibodies coupled to magnetic beads. In both cases, we specifically want to isolate Hsp coupled with other proteins and use the Hsp complexes as intermediaries to present the coupled peptides/proteins to the immune system, or to explore the associations of a particular Hsp with other proteins.

Posible quimera M3: M5 en un paciente con leucemia aguda promielocítica: diferente respuesta clonal al tratamiento con ANTRA y quimioterapia / Probable chimera M3:M5 in an acute promyelocytic leukemia patient: different clonal response to treatment with transretinoic acid and chemotherapy

Se presenta un caso con leucemia aguda promielocítica (LAP) en el que se demostró la presencia del gen híbrido PML/RARa y la duplicación interna en tandem del gen FLT3 (DIT/FLT3) al momento del diagnóstico. Después de recibir tratamiento de inducción con ácido transretinoico (ATRA) y quimioterapia, el estudio citomorfológico de la médula ósea mostró una transformación a leucemia monocítica aguda (LMA-M5). En el estudio molecular desapareció el transcripto PML/RARa, pero se mantuvo la DIT/FLT3. Estos resultados sugieren la coexistencia de 2 clones leucémicos independientes, un clon promielocítico (M3) con el gen quimérico PML/RARa y otro monocítico (M5) con DIT/FLT3. Aunque la evolución hematológica y molecular apoya esta sugerencia, no se puede excluir la presencia de la DIT/FLT3 en el clon M3, pues no es un marcador específico de la LMA-M5(AU)

Posible quimera M3: M5 en un paciente con leucemia aguda promielocítica: diferente respuesta clonal al tratamiento con ANTRA y quimioterapia / Probable chimera M3:M5 in an acute promyelocytic leukemia patient: different clonal response to treatment with transretinoic acid and chemotherapy

Se presenta un caso con leucemia aguda promielocítica (LAP) en el que se demostró la presencia del gen híbrido PML/RARa y la duplicación interna en tandem del gen FLT3 (DIT/FLT3) al momento del diagnóstico. Después de recibir tratamiento de inducción con ácido transretinoico (ATRA) y quimioterapia, el estudio citomorfológico de la médula ósea mostró una transformación a leucemia monocítica aguda (LMA-M5). En el estudio molecular desapareció el transcripto PML/RARa, pero se mantuvo la DIT/FLT3. Estos resultados sugieren la coexistencia de 2 clones leucémicos independientes, un clon promielocítico (M3) con el gen quimérico PML/RARa y otro monocítico (M5) con DIT/FLT3. Aunque la evolución hematológica y molecular apoya esta sugerencia, no se puede excluir la presencia de la DIT/FLT3 en el clon M3, pues no es un marcador específico de la LMA-M5

Estudio del reordenamiento molecular de los genes TEL/AML1 en la leucemia linfocítica aguda: resultados preliminares / I study of the molecular reordenamiento of the genes TEL/AML1 in the leukemia sharp linfocítica. Preliminary results

La leucemia linfocítica aguda (LLA) representa aproximadamente el 80 por ciento dentro de las leucemias pediátricas. Recientemente se ha demostrado por biología molecular la existencia de una translocación críptica, la t (12;21) (p12;q22) en la LLA de tipo B, que no se detecta por las técnicas citogenéticas convencionales e involucra los oncogenes TEL y AML1. Esta alteración es actualmente la más común en esta leucemia y se observa aproximadamente en el 25 por ciento de los casos. Diversos investigadores han planteado que dicha translocación identifica a un subgrupo de pacientes con una evolución muy favorable, por lo que se considera un indicador de buen pronóstico. La determinación de la t (12;21) en el estudio de LLA tiene importancia pronóstica, además de servir como marcador para la detección de la enfermedad mínima residual. En nuestro trabajo estandarizamos la técnica de RT-PCR para la detección de la t (12;21) y además analizamos muestras de 20 pacientes pediátricos con LLA tipo B, que en el momento del estudio se encontraban en la fase de diagnóstico inicial o en recaída. En el estudio, 5 de los 20 pacientes mostraron reordenados los genes TEL/AML 1, lo que representa el 25 por ciento de los casos. Esta frecuencia concuerda con lo comunicado en la literatura hasta el momento(AU)

Estudio del reordenamiento molecular de los genes TEL/AML1 en la leucemia linfocítica aguda: Resultados preliminares / Study of the molecular rearrangement of TEL/AML 1 genes in acute lymphocytic leukemia: Preliminary results

La leucemia linfocítica aguda (LLA) representa aproximadamente el 80 % dentro de las leucemias pediátricas. Recientemente se ha demostrado por biología molecular la existencia de una translocación críptica, la t (12;21) (p12;q22) en la LLA de tipo B, que no se detecta por las técnicas citogenéticas convencionales e involucra los oncogenes TEL y AML1. Esta alteración es actualmente la más común en esta leucemia y se observa aproximadamente en el 25 % de los casos. Diversos investigadores han planteado que dicha translocación identifica a un subgrupo de pacientes con una evolución muy favorable, por lo que se considera un indicador de buen pronóstico. La determinación de la t (12;21) en el estudio de LLA tiene importancia pronóstica, además de servir como marcador para la detección de la enfermedad mínima residual. En nuestro trabajo estandarizamos la técnica de RT-PCR para la detección de la t (12;21) y además analizamos muestras de 20 pacientes pediátricos con LLA tipo B, que en el momento del estudio se encontraban en la fase de diagnóstico inicial o en recaída. En el estudio, 5 de los 20 pacientes mostraron reordenados los genes TEL/AML 1, lo que representa el 25 % de los casos. Esta frecuencia concuerda con lo comunicado en la literatura hasta el momento.

Acute lymphocytic leukemia (ALL) represents approximately 80% of the pediatric leukemias.The existance of a cryptic translocation, the t (12;21) (p12;q22) in the type B ALL, which is not detected by the conventional cytogenetic techniques and involve the TEL and AML1 oncogenes, has been recently shown by molecular biology. This alteration is at present the most common in this leukemia and it is observed approximately in 25% of the cases. Some authors have stated that such translocation identifies a group of patients with a very favorable evolution and that's why it is considered as an indicator of good prognosis. The determination of the t (12;21) in the study of ALL has a prognostic importance and it also serves as a marker for the detection of the minimal residual disease. In our paper, we standardized the RT-PCR technique for the detection of the t (12;21) and we also analized samples from 20 pediatric patients with type B ALL, which at the time of the study was in the phase of initial diagnosis or on relapse. In the study, 5 of the 20 patients showed rearranged TEL/AML l genes, which accounted for 25% of the cases. This frequency agrees with what is reported in literature up to now.

Diagnóstico molecular de la leucemia aguda promielocítica: Resultados preliminares / Molecular diagnosis of the leukemia sharp promielocítica: Preliminary results

La leucemia aguda promielocítica (LAP) se caracteriza por la presencia de la translocación recíproca t (15;17) que tiene como resultado la formación del gen híbrido PML-RARa . Como la LAP se considera una emergencia hematológica y además tiene hoy en día un tratamiento muy específico con ácido retinoico (ATRA), es muy importante hacer un diagnóstico rápido y preciso, que en muchos casos permite incluso salvar la vida del paciente. En la actualidad se han desarrollado métodos de RT-PCR para detectar el gen híbrido PML-RARa . Estas técnicas moleculares han sido extremadamente útiles en el diagnóstico de esta entidad. En este trabajo presentamos los resultados preliminares del diagnóstico molecular en 38 pacientes con LAP. En 36 pacientes se demostró la presencia del gen híbrido y 2 fueron negativos. Del total de enfermos con resultados positivos, 19 (55 por ciento) fueron bcr 1, 2 (5 por ciento) fueron bcr 2 y 14 (40 por ciento) fueron bcr 3. Todos los pacientes con resultados positivos respondieron al tratamiento con ATRA. En 1 de los 2 pacientes con resultados negativos se demostró la presencia de la t(11;17). Ninguno de estos 2 enfermos respondió al tratamiento con ATRA(AU)

Diagnóstico molecular de la leucemia aguda promielocítica: Resultados preliminares / Molecular diagnosis of acute promyelocytic leukemia: preliminary results

La leucemia aguda promielocítica (LAP) se caracteriza por la presencia de la translocación recíproca t (15;17) que tiene como resultado la formación del gen híbrido PML-RARa . Como la LAP se considera una emergencia hematológica y además tiene hoy en día un tratamiento muy específico con ácido retinoico (ATRA), es muy importante hacer un diagnóstico rápido y preciso, que en muchos casos permite incluso salvar la vida del paciente. En la actualidad se han desarrollado métodos de RT-PCR para detectar el gen híbrido PML-RARa . Estas técnicas moleculares han sido extremadamente útiles en el diagnóstico de esta entidad. En este trabajo presentamos los resultados preliminares del diagnóstico molecular en 38 pacientes con LAP. En 36 pacientes se demostró la presencia del gen híbrido y 2 fueron negativos. Del total de enfermos con resultados positivos, 19 (55 %) fueron bcr 1, 2 (5 %) fueron bcr 2 y 14 (40 %) fueron bcr 3. Todos los pacientes con resultados positivos respondieron al tratamiento con ATRA. En 1 de los 2 pacientes con resultados negativos se demostró la presencia de la t(11;17). Ninguno de estos 2 enfermos respondió al tratamiento con ATRA.

Acute promyelocytic leukemia (APL) is characterized by the reciprocal translocation t(15; 17) that results in fusion gene PML-RARa formation. As APL is considered to be an hematological emergency and also is given a very specific treatment with retinoic acid (ATRA), then it is very important to diagnose quickly and accurately which makes it possible in many cases to save the patient's life. At present RT- PCR methods have been developed to detect PML-RARa gen. These molecular techniques have been extremely useful in diagnosing this entity. This paper sets forth the preliminary results of molecular diagnoses of APL in 38 patients. 36 cases presented the fusion gene and 2 did not. Of the total number of positive patients, 19(55%) were bcr 1, 2 (5%) were bcr 2 and 14(40%) bcr 3. All the patients with positive results were responsive to treatment with ATRA. One of the two patients with negative results showed the existence of t (11; 17). These two patients did not respond to treatment with ATRA.

Actividad biológica de la prolactina en pacientes con galactorrea y anovulación / Biological activity of prolactin in patients with galactorrhoea and anovulation

Uno de los efectos más documentados para la prolactina es su actividad lactótrofa y sobre el desarrollo mamario en mamíferos. Existen numerosos casos de pacientes normoprolactinémicas que cursan con galactorrea, en las cuales se desconoce el mecanismo responsable de este signo clínico, postulándose la posible existencia de formas hiperactivas de la hormona. En el presente trabajo se estudió la actividad biológica de la prolactina en pacientes normoprolactinémicas e hiperprolactinémicas con galactorrea y/o anovulación asociada, midiendo la actividad mitogénica de la hormona en un bioensayo de células Nb2 en cultivo, a un grupo de muestras de suero obtenidas en el Laboratorio de Reproducción Asistencial del Instituto Nacional de Endocrinología a las cuales previamente se les determinó el nivel de prolactina sérica inmunorreactiva mediante radioinmunoanálisis; se realizó además un ensayo de inhibición de la actividad biológica de la hormona en presencia de anticuerpos antiprolactina, para medir hasta qué punto la actividad mitogénica detectada se debía a la prolactina presente en las muestras. Los resultados obtenidos demuestran que la galactorrea es independiente de los niveles séricos de prolactina inmunorreactiva, no se demostró la existencia de formas hiperactivas responsables de este signo clínico en los diferentes grupos de pacientes normoprolactinémicas y se corroboró el papel biológico de la hormona en el fallo ovárico que tiene lugar en las pacientes hiperprolactinémicas (AU)

Actividad biológica de la prolactina en pacientes con galactorrea y anovulación / Biological activity of prolactin in patients with galactorrhoea and anovulation

Uno de los efectos más documentados para la prolactina es su actividad lactótrofa y sobre el desarrollo mamario en mamíferos. Existen numerosos casos de pacientes normoprolactinémicas que cursan con galactorrea, en las cuales se desconoce el mecanismo responsable de este signo clínico, postulándose la posible existencia de formas hiperactivas de la hormona. En el presente trabajo se estudió la actividad biológica de la prolactina en pacientes normoprolactinémicas e hiperprolactinémicas con galactorrea y/o anovulación asociada, midiendo la actividad mitogénica de la hormona en un bioensayo de células Nb2 en cultivo, a un grupo de muestras de suero obtenidas en el Laboratorio de Reproducción Asistencial del Instituto Nacional de Endocrinología a las cuales previamente se les determinó el nivel de prolactina sérica inmunorreactiva mediante radioinmunoanálisis; se realizó además un ensayo de inhibición de la actividad biológica de la hormona en presencia de anticuerpos antiprolactina, para medir hasta qué punto la actividad mitogénica detectada se debía a la prolactina presente en las muestras. Los resultados obtenidos demuestran que la galactorrea es independiente de los niveles séricos de prolactina inmunorreactiva, no se demostró la existencia de formas hiperactivas responsables de este signo clínico en los diferentes grupos de pacientes normoprolactinémicas y se corroboró el papel biológico de la hormona en el fallo ovárico que tiene lugar en las pacientes hiperprolactinémicas(AU)