Steroid Receptor Expression Research Articles

Dihydrotestosterone promotes kidney cancer cell proliferation by activating the STAT5 pathway via androgen and glucocorticoid receptors.

Androgen receptors (ARs) are expressed on a variety of cell types, and AR signaling plays an important role in tumor development and progression in several cancers. This in vitro study evaluated the effect of dihydrotestosterone (DHT) on the proliferation of renal cell carcinoma (RCC) cells in relation to AR status. Steroid hormone receptor expression was evaluated using RT-PCR and Western blotting. The effect of DHT on cell proliferation and STAT5 phosphorylation was evaluated in RCC cell lines (Caki-2, A498, and SN12C) and primary RCC cells using cell viability assays and Western blotting. ARs and glucocorticoid receptors (GRs) were knocked down with small interfering RNAs before assessing changes in cell proliferation and STAT5 activation. DHT treatment promoted cell proliferation and increased STAT5 phosphorylation regardless of AR status. The AR antagonist bicalutamide reduced kidney cancer cell proliferation, regardless of AR status. AR and GR knockdown blocked STAT5 activation and reduced cell proliferation in all RCC cell lines. In patient-derived primary cells, DHT enhanced cell proliferation and this effect was diminished by treatment with the AR antagonists bicalutamide and enzalutamide and the GR antagonist mifepristone. DHT promotes cell proliferation through STAT5 activation in RCC cells, regardless of AR status. DHT appears to utilize the AR and GR pathways to activate STAT5, and the inhibition of AR and GR showed antitumor activity in RCC cells. These data suggest that targeting AR and GR may be a promising new approach to the treatment of RCC.

Effects of l-arginine on endometrial estrogen receptor α/β and progesterone receptor expression in nutrient-restricted sheep

This study aimed to determine the effects of l-arginine (L-Arg) supplementation on steroid hormone receptors in non-pregnant ovine endometrium. All experimental ewes were randomly assigned to either a control group (n = 6), a nutrient-restricted group (n = 6), or an L-Arg supplemented nutrient-restricted group (n = 6). The effects of L-Arg on estrogen receptor α/β (ERα/β) and progesterone receptor (PGR) expression in the ovine endometrium were assessed. Our results showed that levels of ERβ and PGR expression were significantly increased by nutrient restriction, but L-Arg counteracted the effect of nutrient restriction on ERβ and PGR expression (p < 0.05). Also, expression of endometrial ERα was substantially increased (p < 0.05) by L-Arg supplementation. Furthermore, ERα/β and PGR were mainly detected in the endometrial luminal epithelium and glandular epithelium. Therefore, we isolated and identified endometrial epithelial cells (EECs) from sheep. Different concentrations of L-Arg were added to investigate the effects on ERα/β and PGR in EECs. The expression levels of endothelial nitric oxide synthase, ERβ, and PGR were significantly increased in response to low-concentration (200 μmol) L-Arg supplementation, which subsequently decreased with a high concentration (800 μmol) (p < 0.05). Otherwise, ERα expression was remarkably increased at both L-Arg concentrations in EECs (p < 0.05). Overall, the results indicated that L-Arg performed crucial roles in the regulation of ovine steroid hormone receptor expression in the endometrium. The results of this study provide a theoretical basis and technical means for the normal function of endometrium in response to low nutrient levels.

P-129 - Expression of female sex hormone receptors, connective tissue growth factor, and HER2 in gallbladder cancer and adjacent normal tissue

P-129 - Expression of female sex hormone receptors, connective tissue growth factor, and HER2 in gallbladder cancer and adjacent normal tissue

Endometriosis-assEndometriosis-associated ovarian tumors: morphological and immunohistochemical features

Background. In 2016, the World Health Organization published an updated version of the Histological Classification for ovarian tumors presenting a new category of endometriosis-associated tumors. The predictors of malignant transformation of endometriosis have not been clearly defined so far.Purpose. The search for histological and immunohistochemical markers of endometriosis-associated malignancy.Materials and methods. 28 female patients with endometrioid ovarian cancer and 11 patients with clear cell ovarian carcinoma were enrolled. Histological and immunohistochemical studies were carried out using conventional techniques. Immunohistochemistry was applied to determine the hormone receptor status: expression of steroid hormone receptors, BAF250a (ARID1A), PTEN, P-catenin, MSH6, PMS2, р-53, WT-1, proliferative index (Ki-67). Microsatellite instability (MSI) testing was conducted according to the standard protocol.Results. In all cases of ovarian cancer, histological examination showed one of the endometriosis features. Atypical endometriosis was found in 39 % (11 / 28) of endometrioid tumors and in 9% (1/ 11) of clear cell carcinomas. Endometrioid ovarian cancer was found to be ER (74±7,8%) — and PR (67±5,4%) — positive; Ki-67 index was 68,2±3,7 %; loss of BAF250a (ARID1A) expression was observed in 14% (4/ 28), loss of PTEN expression in 29 % (8 / 28), nuclear expression of P-catenin in 32% (9/28) of cases. Loss of MMR expression was detected in 7% (2/28) of cases. MSI was found in one case only, which was also associated with loss of expression of BAF250a (ARID1A) and MSH6. Clear cell carcinoma of the ovary showed histological criteria for endometriosis; however, there were no changes immunohistochemical markers expression that were typical for endometriosis-associated malignancies. It could be due to a small number of patients in the group so further research is needed.Conclusion. Atypical endometriosis may be a morphological precursor of endometrioid and clear cell carcinoma of the ovary. Comprehensive assessment of a marker panel consisting of BAF250a (ARID1A), P-catenin, PTEN, p53, Ki-67 index, PMS2 and MSH6 will allow improving the diagnosis of atypical endometriosis and endometriosis-associated ovarian cancer.

Receptor Status of Tumor Cells and Mechanisms of Disorders of Tissue Homeostasis in Carcinogenesis of Serous Ovarian Cancer

The aim of the study is to define the value of sex hormone receptors expression level in serous ovarian cancer tumor cells for different variants of genetic instability of the cellular substrate and to evaluate its pathogenetic role in the biological of the tumor. Material and methods . The study of surgical and biopsy material from 89 patients with serous ovarian cancer was performed in histological preparations stained with hematoxylin and eosin and by the Felgen method. Subsequently, the degree of histological differentiation, mitotic regime and ploidy of tumor cells was evaluated, as well as immunohistochemical analysis of the expression in tumor cells of the tumor suppressor mtp53, progesterone and estrogen receptors. Two study groups were distinguished. The first group consisted of 62.9% of patients with negative p53 IGH status. The second group included 37.1% of cases with positive IGH expression of p53 in cells of serous ovarian cancer. Results. The first group of the studied tumors was characterized by significant morphological heterogeneity. On the contrary, in the group of serous carcinomas where a pronounced expression of oncogen p53 was diagnosed with a pronounced tendency to a low degree of histological differentiation of the parenchymal component of the tumor showed much greater morphological similarity and uniformity. At the same time, tumors in this group were characterized by the maximum indicators of ploidy of tumor cells, which, in our opinion, indicates an increase in cellular atypism and pronounced genetic instability against the background of high proliferative activity. With a significantly different level of genetic instability between the carcinomas of the study groups, the expression of estrogen receptors showed an inverse dependence on the level of cell proliferative activity and the amount of genetic material in the cancer cell. A negative correlation of the level of expression of sex hormone receptors with the degree of tumor differentiation regardless of the level of genetic abnormalities was revealed. Conclusion. It has been shown that the potential of tumor progression and the clinically more aggressive behavior of a neoplastic tissue substrate largely depends on the level of genetic instability in the epithelium of a cancer tumor, but at the same time, the altered regulatory mechanisms of cellular interactions (dysfunction of paracrine and endocrine regulation) in the tumor type cells leading to a violation of tissue homeostasis.

The Notch Family Transcription Factor, RBPJκ, Modulates Glucose Transporter and Ovarian Steroid Hormone Receptor Expression During Decidualization.

During decidualization, endometrial stromal cells differentiate into a secretory phenotype to modulate the uterine microenvironment and promote embryo implantation. This highly metabolic process relies on ovarian steroid receptors and glucose transporters. Canonical Notch signaling is mediated by the transcription factor Recombination Signal Binding Protein for Immunoglobulin Kappa J Region (RBPJ). Loss of RBPJ in the mouse uterus (Pgrcre/+Rbpjflox/flox; Rbpj c-KO) results in subfertility in part due to an abnormal uterine-embryonic axis during implantation and, as described herein, decidualization failure. Induced in vivo decidualization in Rbpj c-KO mice was impaired with the downregulation of decidual markers and decreased progesterone receptor (Pgr) signaling. Consistent with in vivo mouse data, RBPJ knockdown during in vitro Human uterine fibroblast (HuF) cell decidualization results in the reduced expression of decidual marker genes along with PGR. Expression of the glucose transporter, SLC2A1, was decreased in the RBPJ-silenced HuF cells, which corresponded to decreased Slc2a1 in the secondary decidual zone of Rbpj c-KO mouse uteri. Exogenous administration of pyruvate, which bypasses the need for glucose, rescues PRL expression in RBPJ-deficient HuF cells. In summary, Notch signaling through RBPJ controls both ovarian steroid receptor PGR and glucose transporter SLC2A1 expression during decidualization, and this dysregulation likely contributes to embryo implantation failure.

Aberrant endometrial steroid receptor expression in in-vitro maturation cycles despite hormonal luteal support: A pilot study

Clinical outcomes of fresh embryo transfer in non-hCG triggered in vitro maturation (IVM) cycles are inferior compared to vitrified-warmed embryo transfer. This is a prospective observational pilot study in a consecutive cohort of 31 polycystic ovary syndrome (PCOS) patients and 37 normo-ovulatory egg donors who underwent IVM without fresh embryo transfer between July 2009 and June 2014. All subjects received 150 IU of highly purified menotropin (HP-hMG) daily for three days. On cycle day 6, all patients started transdermal oestradiol (E2) at a daily dose of 9 mg. There was no human chorionic gonadotropin (hCG) trigger before oocyte retrieval (OR). Vaginal micronized progesterone was commenced on the evening after OR, at a daily dose of 600 mg. Additional luteal phase support (LPS) was administered as follows: Group A: no additional LPS; Group B: 1500 IU of hCG administered 4 h after OR and Group C: 5000 IU of hCG administered 4 h after OR + an additional injection of 5000 IU of hCG 1 day before endometrial biopsy. Endometrial biopsy for histology and immunohistochemistry (IHC) was performed on day 5 or 6 after OR. Instead of being downregulated, both PR-B and ERα in endometrial glands and stroma were moderately to strongly expressed in all three protocols, suggesting that the mid-luteal histological signature of endometrial receptivity is deficient in a non-hCG-triggered IVM cycle. Poor clinical outcomes after fresh embryo transfer following IVM are probably related to inappropriate endometrial development which may be linked to the short follicular phase of IVM cycles.

Steroidogenic potential of human fetal kidney at early gestational age

Steroidogenic potential of the human fetal kidney (hFK) at the end of first trimester is poorly investigated. Little is known about the ontogeny of steroidogenic enzymes and activities of steroidogenic pathways in the hFK at early pregnancy.Our aim was to explore steroidogenesis and the expression of steroidogenic enzymes in the hFK at gestational weeks (GW) 9-12. Steroids in the hFK were analyzed by gas chromatography/coupled to tandem mass spectrometry. The expression of steroidogenic enzymes in the hFK at GW 9-12 was investigated by qPCR, automated Western blotting and immunohistochemistry. We observed that the hFK produced substantial amount of steroids of the Δ5 and Δ4 pathways and several steroid precursors in the biosynthesis of DHT via the backdoor pathway but not DHT itself. The levels of steroids and expression of relevant steroidogenic enzymes (e.g., CYP17A1, HSD3B1, HSD3B2, CYP11B1 and AKR1C4) we significantly higher in the hFK at GW11-12 compared to GW9. We also found the expression of sex steroid receptors (e.g., AR, ERα and ERβ) in the hFK at GW9-12. No sex-dependent differences in the levels of all identified steroids and expression of steroidogenic enzymes in the hFK from male and female fetuses were found.Altogether, our data indicate that the hFK at early pregnancy is steroidogenic organ with potential to synthesize multiple steroids that may play an important role in the formation and development of this organ in humans.

Prevalence of PgR, ER and HER2+ receptors among women with breast cancer by age in Poland

Introduction. Prevalence of estrogen (ER), progesterone (PgR) and human epidermal growth factor receptors (HER2) vary by age among women with breast cancer (BC). Such variation has a large significance for the prognosis and treatment process. This study characterizes the prevalence of breast cancer subtypes by age in a hospital sample in Poland. Material and methods. The study population included 735 women with BC aged 27–91 years old (ICD-10: C50) and treated in the years 2009–2011 in the Military Institute of Medicine in Warsaw. Subjects were divided into 2 age categories — 27–44 and 45+ — which included 66 (9%) and 669 (91%) women respectively. In each group prevalence of PgR, ER and HER2+ receptors was investigated. Results. In both age groups the most frequent BC subtype was luminal A (hormone dependent BC — with PgR and ER receptor expression) — 27–44 years old — 44% and 45+ years old — 56%. The lowest number of cases in the age group 27–44 was luminal B (triple positive breast cancer) — about 15% of cases and for 45+ age group — HER2+ BC — about 11%. Discussion. Performed research shows relationship between growing age of diagnosis and presence of more desirable features of BC among women aged 55 and more, such as expression of steroid receptors or lack of expression of HER2 receptors, which is a good prognostic indicator for treatment outcomes. In the same time, many studies suggest that more aggressive types of breast cancer (basal-like — triple negative) are more characteristic for younger age groups (under 45 y.o. and younger women in perimenopausal age). Same results have been obtained in own study. Conclusions. A high incidence of BC in older age groups (45+) and more frequent occurrence of aggressive types of BC among younger women (27–44 years old) indicate the need to educate women from both age groups about risk factors and early symptoms of the disease. As we still have not recognized all BC risk factors, education about well-known agents, such as alcohol intake, overweight and obesity, play significant role in decrease of BC incidence and mortality.

Effects of Metformin on Cellular Proliferation and Steroid Hormone Receptors in Patient-Derived, Low-Grade Endometrial Cancer Cell Lines.

Endometrial cancer (EC) is the most common gynecologic malignancy and is the result of disruption of the balance between estrogen-stimulated growth and progesterone-induced growth modulation. Metformin has been shown to inhibit EC proliferation; however, its role in early-stage EC and its effects on steroid hormone receptors have not been adequately explored. Our aim was to examine the effects of metformin on cellular proliferation in patient-derived, low-grade EC cell lines and to determine whether it directly modulates steroid hormone receptor expression. Two novel EC cell lines were produced (EM2 and 3) from endometrial tumor tissue obtained from women undergoing surgery. Cellular proliferation was determined by the 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay, and in both cell lines, metformin decreased cell proliferation in a dose-dependent (10-200 µmol/L) manner and induced apoptosis as measured by cleaved PARP. Furthermore, metformin abrogated the effects of E2 on cell proliferation. Using quantitative real-time polymerase chain reaction and Western immunoblotting, metformin significantly decreased estrogen receptor (ER) α messenger RNA abundance but did not consistently affect the expression of progesterone receptor. Estrogen receptor α protein levels significantly decreased across all metformin doses tested, which resulted in a significant decrease in the expression of the ER targets genes Keratin-19 and Wnt-1 inducible signaling pathway 2. In addition, metformin increased phosphorylation of AMPK in a dose-dependent manner (10-200 µmol/L) indicating an effect on mammalian target of rapamycin (mTOR) signaling. Our data suggest that metformin therapy represents a potential fertility-sparing option for women with early-stage EC, given its capacity to inhibit EC cell proliferation, ERα expression, and the mTOR cell proliferation pathway.

SUN-352 Estrogen, Progesterone, and Androgen Receptor Expressions Predict the Prognosis for Adrenocortical Carcinoma

Aim: Adrenocortical carcinoma (ACC) is a rare but mortal endocrine malignancy. While expression of sex steroid receptors is well-known to be associated with tumor prognosis and treatment response in a number of hormone sensitive cancers, sex steroid receptor expression in ACC is not revealed in this context. The aim of this study is to assess prognostic implications of sex steroid receptor expression in ACCs. Method: Paraffin-embedded blocks of 16 ACC cases (8 F, 8 M, mean age 47.9±13.9) and 22 benign adrenocortical adenoma (ACA) cases (14 F, 8 M, mean age 50.0±11.4) who were undergone surgery in our center between 1998-2017 were examined in this case-control study. Estrogen (ER, Leica), androgen (AR, CellMarque) and progesterone receptor (PR, Leica) expressions in both tumor and peritumoral adrenal tissue were semi-quantitatively scored by immunohistochemistry (IHC), by an endocrine pathology specialist. The relationship between the histopathological prognostic indicators, the clinical course of patients and intra-tumor and peritumoral sex steroid receptor expression status were examined. Results: ER and AR expressions of ACCs were significantly higher, while all ACAs were ER and AR negative (ER 25% vs 0%, p=0.014; AR 31% vs 0%, p=0.005). PR expression was similar between groups; although, higher rate of PR expression was observed in ACAs (75% vs 86%, p=0.38). While peri-tumoral adrenal tissue of ACC cases did not express none of the three sex steroid receptors, IHC staining of peripheral adrenal tissue of ACAs revealed positivity for ER, AR and PR (p values <0.005). ACC cases which were negative for ER, PR and/or AR had larger tumor size (r=-0.6, p=0.015). Tumor stage was more advanced in AR negative ACCs (r=-0.6, p=0.021). Progression free survival was shorter in ACC cases which were positive for PR (15 vs 57.5 months, p=0.07); while it was longer in cases peritumoral adrenal tissue was positive for either of three sex steroid receptors (r=0.81, p=0.008). Conclusions: For the first time in this study, expression of sex steroid receptors has been shown to be related with ACC prognosis and clinical/histopathological behaviour.

SUN-021 Chronic Psychosocial Stress during Pregnancy Affects Maternal Behavior and Neuroendocrine Function and Modulates Hypothalamic CRH Signaling Pathway and Nuclear Steroid Hormone Receptor Expression

Postpartum depression (PPD) affects up to 20% of women and exerts adverse consequences on mother and child. Gestational stress and abnormalities in neuroendocrine function have been implicated in the development of PPD. Here, we measured effects of chronic psychosocial stress during pregnancy on maternal behavior as well as hypothalamic-pituitary-adrenal (HPA) axis function and regulation in the early postpartum period. From gestational day 6.5 to 17.5, pregnant C57Bl/6 female mice were exposed to a novel chronic stress paradigm consisting of variable psychosocial stressors such as foreign object exposure, rat odor exposure, bedding removal and were assessed from postpartum day 2 to 7 for behavioral alterations in maternal care, depression, and anxiety as well as circadian and brief restraint-stress associated corticosterone response. mRNA changes in molecular regulators of the HPA axis were measured in 1mm micropunches of the hypothalamic paraventricular nucleus (PVN) via qPCR. Mice exhibited deficits in maternal care after undergoing chronic psychosocial stress during pregnancy displayed as increased latency to retrieve pups during pup retrieval task (p<0.01, n=17). Furthermore, they displayed a depressive-like phenotype as measured by increased time spent immobile in forced swim test (p<0.01, n=17) and increased anxiety as measured by increased time spent in dark zone in light/dark transition test (p<0.05, n=7-11). Abnormalities in the activity of the maternal HPA axis were also displayed as seen by a flattening of the circadian rhythm of CORT secretion (p<0.05, n=10-12), increased CORT release following 20 minutes of restraint stress (p<0.05, n=5-8), and increased adrenal gland weights (p<0.05, n=15-20). These changes were associated with increased PVN CRH mRNA (gestational day 17.5 (G17.5): 0.880+0.18 vs. 0.485+0.03, postpartum day 2 (PP2): 1.012+0.05 vs. 0.794+0.05, PP7: 0.775+0.04 vs. 0.487+0.05, p<0.05, n=6-7) and downregulation of CRH receptor 1 (G17.5: 0.821+0.07 vs. 1.03+0.05, PP2: 1.007+0.05 vs. 1.184+0.06, p<0.05, n=6-7) compared to non-stressed control dams, while vasopressin and oxytocin levels remained undisturbed. Furthermore, PVN glucocorticoid receptor (G17.5: 0.844+0.01 vs. 0.938+0.02, p<0.01, n=6-7) and progesterone receptor mRNA were downregulated (PP2: 1.008+0.03 vs. 1.196+0.047, p<0.01, n=6-7), and FK506 binding protein 5 mRNA, a co-chaperone known to negatively orchestrate GR/PR transcriptional activity, was upregulated (G17.5: 2.322+0.37 vs. 1.315+0.05, p<0.05, n=6-7). These results suggest hypothalamic changes in GR/PR signaling pathway as putative regulators of postpartum changes in CRH after stress exposure and postpartum maternal affective dysregulation. The mechanistic role GR/PR play in mediating these changes is further being investigated by selective spatiotemporal GR/PR modulation using Cre-loxP technology.

Low Daytime Light Intensity Disrupts Male Copulatory Behavior, and Upregulates Medial Preoptic Area Steroid Hormone and Dopamine Receptor Expression, in a Diurnal Rodent Model of Seasonal Affective Disorder.

Seasonal affective disorder (SAD) involves a number of psychological and behavioral impairments that emerge during the low daytime light intensity associated with winter, but which remit during the high daytime light intensity associated with summer. One symptom frequently reported by SAD patients is reduced sexual interest and activity, but the endocrine and neural bases of this particular impairment during low daylight intensity is unknown. Using a diurnal laboratory rodent, the Nile grass rat (Arvicanthis niloticus), we determined how chronic housing under a 12:12 h day/night cycle involving dim low-intensity daylight (50 lux) or bright high-intensity daylight (1,000 lux) affects males’ copulatory behavior, reproductive organ weight, and circulating testosterone. We also examined the expression of mRNAs for the aromatase enzyme, estrogen receptor 1 (ESR1), and androgen receptor (AR) in the medial preoptic area (mPOA; brain site involved in the sensory and hormonal control of copulation), and mRNAs for the dopamine (DA) D1 and D2 receptors in both the mPOA and nucleus accumbens (NAC; brain site involved in stimulus salience and motivation to respond to reward). Compared to male grass rats housed in high-intensity daylight, males in low-intensity daylight displayed fewer mounts and intromissions when interacting with females, but the groups did not differ in their testes or seminal vesicle weights, or in their circulating levels of testosterone. Males in low-intensity daylight unexpectedly had higher ESR1, AR and D1 receptor mRNA in the mPOA, but did not differ from high-intensity daylight males in D1 or D2 mRNA expression in the NAC. Reminiscent of humans with SAD, dim winter-like daylight intensity impairs aspects of sexual behavior in a male diurnal rodent. This effect is not due to reduced circulating testosterone and is associated with upregulation of mPOA steroid and DA receptors that may help maintain some sexual motivation and behavior under winter-like lighting conditions.

Sex steroid hormone receptor expression in the vaginal wall in cervical cancer survivors after radiotherapy

Background: Sex steroid hormones and their receptors are important in female sexual function. The aim of this study was to investigate the expression and distribution of estrogen receptor (ER)α, ERβ, G-protein-coupled ER-1 (GPER), androgen receptor (AR), progesterone receptor (PR)A, PRB and connective tissue growth factor (CTGF) in the vaginal wall among women who had been treated for cervical cancer with radiotherapy.Material and methods: We included cervical cancer survivors treated with radiotherapy and premenopausal control women of the same age scheduled for benign gynecological surgery. We analyzed the expression and distribution of sex steroid hormone receptors and CTGF in biopsies from the vaginal wall, by real-time PCR and immunohistochemistry (IHC). Serum samples were analyzed for hormone levels and radiation dose at biopsy site were calculated and correlated to levels of the sex steroid hormone receptors.Results: In the cervical cancer survivors (n = 34), we found a lower expression of ERα at both mRNA and protein levels, compared to the control women (n = 37). In the survivors with high radiation dose at biopsy site, the immunostaining of ERα and AR was lower in the epithelium and the stroma, compared to survivors with minimal radiation dose. The later group showed expression of ERα comparable to the control women. The cancer survivors were sufficiently substituted with systemic estradiol with no difference in the serum estradiol levels compared to control women.Conclusions: We found that external radiation reduces the ERα and AR protein expression in the vaginal mucosa, indicating that the vaginal changes in irradiated cervical cancer survivors and the lack of response to hormonal treatment could be due to the decreases in sex steroid hormone receptor expression.

Disorders affecting vitamin B6 metabolism.

Vitamin B6 is present in our diet in many forms, however, only pyridoxal 5'-phosphate (PLP) can function as a cofactor for enzymes. The intestine absorbs nonphosphorylated B6 vitamers, which are converted by specific enzymes to the active PLP form. The role of PLP is enabled by its reactive aldehyde group. Pathways reliant on PLP include amino acid and neurotransmitter metabolism, folate and 1-carbon metabolism, protein and polyamine synthesis, carbohydrate and lipid metabolism, mitochondrial function and erythropoiesis. Besides the role of PLP as a cofactor B6 vitamers also play other cellular roles, for example, as antioxidants, modifying expression and action of steroid hormone receptors, affecting immune function, as chaperones and as an antagonist of Adenosine-5'-triphosphate (ATP) at P2 purinoceptors. Because of the vital role of PLP in neurotransmitter metabolism, particularly synthesis of the inhibitory transmitter γ-aminobutyric acid, it is not surprising that various inborn errors leading to PLP deficiency manifest as B6 -responsive epilepsy, usually of early onset. This includes pyridox(am)ine phosphate oxidase deficiency (a disorder affecting PLP synthesis and recycling), disorders affecting PLP import into the brain (hypophosphatasia and glycosylphosphatidylinositol anchor synthesis defects), a disorder of an intracellular PLP-binding protein (PLPBP, previously named PROSC) and disorders where metabolites accumulate that inactivate PLP, for example, ALDH7A1 deficiency and hyperprolinaemia type II. Patients with these disorders can show rapid control of seizures in response to either pyridoxine and/or PLP with a lifelong dependency on supraphysiological vitamin B6 supply. The clinical and biochemical features of disorders leading to B6 -responsive seizures and the treatment of these disorders are described in this review.

Effects of acute and chronic zinc exposure on steroid hormone receptor expression in the lined seahorse, Hippocampus erectus

The element zinc (Zn) can elicit a variety of physiological responses in animals, particularly through its effects on steroid hormone receptor expression. Here, we evaluated the effects of Zn on the expression profiles of steroid hormone receptors in the lined seahorse, Hippocampus erectus, following experimental acute and chronic Zn exposure. Complex response patterns were observed; however, steroid hormone receptor expression in liver and kidney following 12‐day Zn injection (acute exposure) differed significantly from the control treatments. Apparent effects on hepatic lipid metabolism and osmotic regulation in the kidney need detailed investigation. Furthermore, 49‐day (chronic) waterborne Zn exposure produced a significant decrease in both estrogen receptor (ER) expression levels and estradiol concentrations. In additional, chronic exposure led to a substantial reduction of hepatic adipose cells, implying that Zn may regulate hepatic lipid metabolism via ER and estradiol. Our study demonstrated the effects of both acute and chronic Zn exposure on expression patterns of steroid hormone receptors, which will contribute to understanding the mechanism of Zn‐induced hepatic lipid metabolism and other physiological responses in seahorses.

Expression of Lin28 is correlated with prognosis and expression of HER-2 and steroid receptors in breast cancer.

ObjectiveCumulative data from clinical trials suggest that Lin28 may contribute to poor survival in breast cancer patients. The purpose of this study was to investigate the relationship between Lin28 expression and breast cancer patients’ clinicopathological parameters.MethodsData from a total of 291 breast cancer patients were collected in this study. The expression level of Lin28 was assessed by immunohistochemical staining. The correlation of Lin28 expression and clinicopathological parameters was statically evaluated and the prognostic significance of Lin28 expression was assessed by univariate and multivariate analyses.ResultsOne hundred and eight out of 291 (37.1%) breast cancer specimens showed Lin28 protein positive expression, while the remaining 183 specimens showed negative expression. Positive expression of Lin28 was associated with lymph node metastases (P<0.001), HER-2 (P=0.024), estrogen receptor (P=0.039), and progesterone receptor (P=0.027). Kaplan–Meier analysis showed that Lin28 positive expression showed lower overall survival rates compared with Lin28 negative patients (P=0.019). In the multivariate analysis, Lin28 remained a significant independent prognostic factor (P=0.038) for overall survival rates.ConclusionLin28 expression was associated with advanced disease stage and subtype in breast cancer patients, and Lin28 expression may serve as an independent prognostic factor. These data indicate that Lin28 may play a major role in the therapeutic management of breast cancer.

RET in breast cancer: pathogenic implications and mechanisms of drug resistance.

Initiation, progression, outcome and sensibility to therapies in breast cancer (BC), the most frequent cancer in women, are driven by somatic and germline mutations. Although the effectiveness of hormonal therapies is well-founded, it is prescribed for cancers which express steroid hormone receptors, such as estrogen receptor (ER). RET is a proto-oncogene encoding a transmembrane tyrosine kinase receptor that is activated by one of its four ligands (GDNF, neurturin, artemin or persephin) and one of its coreceptors (Gfrα1-4). Loss-of-function mutations in RET are responsible for Hirschsprung disease, while gain-of-function mutations for multiple endocrine neoplasia type 2. In addition, deregulation of its intracellular signaling, due to mutations, gene rearrangements, overexpression or transcriptional upregulation, can cause several neuroendocrine and epithelial tumors. In BC, amplification of receptor tyrosine kinases, such as ERBB2, EGFR, IGFR and FGFR1, and/or their upregulation contribute to cancer initiation and progression. RET can also have an important role in BC, but only in the subset of ER-positive (ER+) tumors, where it is found overexpressed. Targeting the RET pathway and shedding light on molecular basis of the resistance to hormone therapy may lead to new therapies in ER+ BC, improving treatment outcome and preventing tumor-related events. Thus, here, we review the state of the art of RET biology in BC and agents targeting RET tested in the clinical trials and discuss the specificity of the still available RET inhibitors and the molecular mechanisms underlying the BC resistance to endocrine therapy.

The influence of the expression of steroid receptors on angiogenesis, proliferation and apoptosis in myomas of pre- and postmenopausal women

Introduction/Objective. The aim of this study was to determine the effects of the estrogen and progesterone receptor status on angiogenesis, proliferation, and apoptosis of myoma cells in premenopausal (PreM) and postmenopausal (PostM) women. Methods. This was a cross section; clinical-experimental, retrospective, non-interventional study in the field of the study of fundamental pathogenesis mechanisms of disease using pathohistological materials from the existing archive. The research included 76 patients diagnosed with uterine leiomyomas, operatively treated in the Clinic for Gynecology and Obstetrics, Clinical Centre Kragujevac, Serbia. According to the menstrual status, we formed two experimental subgroups. The first group was PreM women (n = 35; 46.2 ? 5.02 years old), and the second group was PostM women (n = 41; 60.25 ? 5.41 years old). Hematoxylin-eosin staining for myoma and myometrium was conducted, as well as immunohistochemistry for ER?, ER?, PR?, vascular endothelial growth factor, endoglin, Ki67, and caspase-3. Results. Progesterone receptor was overexpressed in myoma and myometrium of PreM compared to myoma and myometrium of PostM women. Expression of caspase-3 was a statistically significant increase in PostM women compared to PreM group. ER? and ER? were not changed among groups neither in myoma nor in myometrium samples. Conclusion. According to our data, PR? had higher influence on apoptosis and cell growth than estrogen receptors. Since PR? was increased in PreM in both myoma and myometrium, probably this expression led further to lower expression of apoptotic marker in PreM women.

Association of anti‐inflammatory and antiangiogenic therapies negatively influences prostate cancer progression in TRAMP mice

Chronic inflammation has been implicated in cancer etiology and angiogenesis is stimulated in this disease. In prostate, the crosstalk between malignant epithelial cells and their microenvironment is an essential step of tumorigenesis during which glandular stroma undergo changes designated as reactive stroma. Thus, the aim herewith was to evaluate the effects of associating anti-inflammatory and antiangiogenic therapies on cancer progression, correlating them with steroid hormone receptor (AR and ERα), reactive stroma (vimentin, αSMA, and TGF-β), and cell proliferation (PCNA) markers expression in the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) model. TRAMP mice (12-week old) were divided into the groups: Control (TRCON): received the vehicles used for drug dilution; Celecoxib (TRCEL): received oral doses of the anti-inflammatory drug celecoxib (15 mg/kg) twice daily; Nintedanib (TRNTB): received oral doses of the antiangiogenic drug nintedanib (10 mg/kg) daily; Nintedanib+Celecoxib (TRNTCEL): received the combination of drugs. After 6 weeks, mice were euthanized and ventral prostate samples were harvested for morphological, immunohistochemical, and Western blotting analyses. While celecoxib led to fibromuscular hypertrophy attenuation, nintedanib significantly reduced the incidence of well-differentiated adenocarcinoma (WDAC) foci in relation to controls, both when administered per se or in association to celecoxib. Furthermore, drug combination was associated with unique effects, including lower incidence of HGPIN lesions; lower AR stromal distribution; changes in ERα localization from epithelial nuclei to stroma as well as significant decrease of TGF-β levels and associated angiogenesis. In parallel, all treatments applied resulted in reduced inflammatory marker and vimentin (VIM) expression. Celecoxib plus nintedanib is an effective antitumor combination against prostate cancer progression in TRAMP mice, showing remarkable efficacy in relation to isolated therapies. Importantly, this efficacy might be due to drug association effect on driving AR and mainly ERα distribution in the prostatic tissue towards benign patterns. In addition, celecoxib and nintedanib impaired the development of a stromal reaction by reducing the recruitment of reactive stroma cells and maintaining a normal smooth muscle cell-rich prostate stroma in TRAMP mice. Collectively, these findings pointed to the beneficial effects of combining anti-inflammatory and antiangiogenic strategies to prevent or delay prostatic tumorigenesis.