miR-200b-3p accelerates progression of pituitary adenomas by negatively regulating expression of RECK

MicroRNA(miR)-200b-3p has been associated with many tumors,but its involvement in pituitary adenoma is unclear.This study investigated the molecular mechanism underlying miR-200b-3p regulation in pituitary adenomas to provide a theoretical basis for treatment.Bioinformatics was used to analyze pituitary adenoma-related genes and screen new targets related to RECK and miRNA.As well,the relationship between miR-200b-3p and RECK protein was verified using a double-luciferase reporter gene assay.The expression of miR-200b-3p in clinical samples was analyzed by in situ hybridization.Transfection of the miR-200b-3p inhibitor and small interfering-RECK(si-RECK)was verified by qPCR.GH3 cell viability and proliferation were detected using CCK8 and EdU assays.Apoptosis was detected by flow cytometry and western blotting.Wound healing and Transwell assays were used to detect cell migration and invasion.The effects of miR-200b-3p and RECK on GH3 cells were verified using salvage experiments.miR-200b-3p was highly expressed in pituitary tumor tissue.Inhibitors of miR-200b-3p inhibited cell proliferation promoted cell apoptosis,inhibited invasion and migration,and inhibited the expression of matrix metalloproteinases.Interestingly,miR-200b-3p negatively regulated RECK.The expression of RECK in pituitary adenoma tissues was lower than that in neighboring tissues.Si-RECK rescued the function of miR-200b-3p inhibitors in the above cellular behaviors,and miR-200b-3p accelerated the development of pituitary adenoma by negatively regulating RECK expression.In summary,this study investigated the molecular mechanism by which miR-200b-3p regulates the progression of pituitary adenoma through the negative regulation of RECK.The findings provide a new target for the treatment of pituitary adenoma.

Profile of Pituitary Adenomas in Senegal: Epidemiological, Clinical and Therapeutic Aspects about 241 Collected Cases

Introduction: in adults, adenomas are the most frequent causes of pituitary tumours. The objective was to characterize its epidemiological and clinical aspects, as well as its therapeutic modalities in Senegal. Methodology: this was a multicenter, retrospective, descriptive and analytical study carried out from the Senegalese register of pituitary adenomas (2008-2022). Results: 242 patients were collected with an average age of 42.4 ± 13 years and a sex ratio (M/F) of 0.91. The mean consultation time was 19.7 ± 23 months. The circumstances of discovery were pituitary apoplexy (11 cases, 4.54%), tumor syndrome (176 cases, 72.72%), gonadal manifestations (103 cases, 42.56%), dysmorphic syndrome (21 cases, 8.68%), hypercorticism (15 cases, 6.19%). Morphologically, it was a macroadenoma (197 cases, 81.40%), an extensive adenoma (22 cases, 9.10%). The hormonal profile was lactotropic hypersecretion (80 cases, 33.05%), somatotropic (13 cases, 5.39%), corticotropic (14 cases, 5.78%), mixed (9 cases, 3.71%). The factors significantly associated with prolactinoma were young age (p = 0.000), female sex (p = 0.000), long consultation delay (p = 0.000) and microadenoma (p = 0.001). Only arterial hypertension was associated with acromegaly (p = 0.000). We found a significant correlation between Cushing’s disease and microadenomas (p = 0.000). Non-secreting adenomas were significantly associated with advanced age (p = 0.000), long delay in consultation (p = 0.000), male gender (p = 0.000), tumor syndrome (p = 0.001) and macroadenomas (p = 0.000). Pituitary surgery was effective in 173 patients (71.49%) including 166 cases (68.59%) by transphenoidal approach. Postoperative incidents were transient diabetes insipidus in 82 cases (47.39%), cerebrospinal fluid leak (20 cases, 11.56%). Death was observed in 18 patients in the entire series (7.44%). The latter had a tumor syndrome (88.88%), a macroadenoma (83.33%) and had undergone pituitary surgery (72.22%). Conclusion: pituitary pathology has become a reality with clinical and secretory polymorphism. The diagnostic delay explains the predominance of macroadenomas which are sources of pre and post-operative complications.

Expression for peroxisome proliferator-activated receptor gamma in pituitary adenomas 38 cases for semi-quantitative immunohistochemical analysis

BACKGROUND： It has been reported that peroxisome proliferator-activated receptor γ （PPAR γ ） is highly expressed in lung cancer, colon cancer, and gastric cancer, as well as other tumors.OBJECTIVE： To study expression of PPAR γ in pituitary adenomas and analyze the role of PPAR γ in hormonal typing of pituitary adenomas. DESIGN, TIME AND SETTING： Semi-quantitative immunohistochemistry of pathological specimens. The experiment was conducted at the Department of Neurosurgery, Wuxi Second Hospital Affiliated to Nanjing Medical University between January 2002 and May 2005. MATERIALS： Surgical resection samples of pituitary adenomas from 38 cases （18 male and 20 female） were analyzed. Eight cases were determined to be invasive pituitary adenomas and 30 cases were non-invasive pituitary adenomas. Hormonal classification of the types of pituitary adenomas revealed somatotrophic adenomas in six cases, corticotrophic adenoma in five cases, prolactinomas in 13 cases, multi-hormone secreting adenomas in six cases, and eight cases of adenoma without altered endocrine function. Five autopsy specimens were collected during the same period from patients of matching age that died from unrelated diseases and were included as normal anterior pituitar3, controls. METHODS： Cell counts for positive immunohistochemical signals were recorded from histopathological sections. The percentage of positive cells was reported as a semi-quantitative analysis. MAIN OUTCOME MEASURES： The rate of PPAR γ positive cells in different types of adenoma was based on hormonal levels and invasiveness of pituitary tumor cells. RESULTS： All tumor biopsies were determined to express PPAR γ. The rate of PPAR γ -positive cells ranged between 8%-65% in the pituitary adenomas. According to hormonal type, PPAR γ expression did not vary between the groups. In addition, there was no significant difference in PPAR γ expression between the non-invasive and invasive pituitary adenomas. CONCLUSIONS： Human pituitary adenomas express PPAR γ, and this expression is unrelated to hormonal type and invasiveness.

Magnetic Resonance Imaging and Biological Markers in Pituitary Adenomas with Invasion of the Cavernous Sinus Space

Objective: To investigate the predictability of MRI and the possiblebiological markers of cavernous sinus invasion of pituitary adenomas associated with fourphenomenas: angiogenesis, cell proliferation, apoptosis and matrix metalloproteinase. Methods: Weevaluated 45 patients with pituitary adenoma according to the MRI, surgical findings and theimmunohistochemistry staining of tumor tissues. Results: The results have shown that the sensitivityof MRI for predicting cavernous sinus invasion in this prospective study was 60%, its specificity85%, its positive predictive value 83.33%, negative predictive value 62.96%. 45 specimens ofpituitary adenomas were analyzed for expression of F8, VEGF, Ki-67, c-myc, Bcl-2, nm23 and MMP-9immunoreactivity using immunoperoxidase staining. MVD was assessed using F8-related antigen. Theresults have shown that MVD of invasive pituitary adenomas was significantly higher than that ofnoninvasive (P 【 0.001). There was an association between the invasion of pituitary adenomas andKi-67 LI (P = 0.039) or the expression of VEGF (P 【 0.001) and MMP-9 (P 【 0.001). But c-myc LI andBcl-2 expression have no association with invasiveness of pituitary adenomas (P = 0.061 versus P =0.201). On the other hand, there is an inverse relationship between nm23 expression and tumorinvasion (P 【 0.001). Conclusion: Parasellar extension of pituitary adenomas through the medial wallof the cavernous sinus is diagnosed at surgery, and with sensitive gadolinium-enhanced MRI, itsextent can be partly determined by radiology. Although our study has shown that MVD and theexpression of VEGF, Ki-67, nm23 and MMP-9 have associations with invasiveness of pituitary adenomas,they are lack of specificity. These markers can only provide some useful information.

IN VITRO CELL CULTURE AND HORMONE RADIOIMMUNOASSAY OF HUMAN PITUITARY ADENOMAS

Tissues from 30 human pituitary adenomas are monolayer-cell-cultured in vitro.Hormone secretion of GH, PRL, TSH, LH and FSH by cells into medium is detected by radioimmunoassay. The pattern and amount of hormone(s) in the medium are used to determine the nature of the cells and thus to establish functional classification of pituitary adenomas. The results show that cell culture technique provides an easy and suitable mode for investigating the nature of pituitary adenomas. Hormone radioimmunoassay of culture medium is precise and reliable and represents the whole adenoma tissue. Further studies can lead to clearer understandings of the pathology of pituitary adenomas.

Comparative Study between Microscopic and Endoscopic Trans-Sphenoidal Surgery for Growth Hormone-Secreting Pituitary Adenomas with Super-Sellar Extension

Background: Advances in the era of modern micro-neurosurgery enabled a reduction of surgical invasiveness and brain retraction which has been defined as minimally invasive or keyhole surgery. Sinonasal endoscopy has brought radical changes in the concepts of pathophysiology and treatment of sinonasal aliments as well as surgical techniques. Aim of the Study: To compare between the use of endoscopic and microscopic trans-sphenoidal approach in resection of growth hormone-secreting pituitary adenomas with Suprasellar Extension. Patients and Methods: This is a prospective study. It had been conducted upon 20 patients having growth hormone secreting pituitary adenoma admitted to Neurosurgery department in Alzar University hospitals and Nasr City Insurance hospital from 2015 to 2018, divided into 2 groups;group A (10 cases) underwent endoscopic endonasal trans-sphenoidal pituitary adenoma resection, while group B (10 cases) operated upon using the standard microscopic trans-sphenoidal pituitary adenoma resection. The inclusion criteria were included: All patients with growth hormone secreting pituitary adenomas showing manifestations of acromegaly, mass effect or hormonal disturbance. Results: This study showed that improvement in outcome was higher in endoscopic group opposed to microscopic group (100% vs. 71% improved headache, 80% vs. 60% visual improvement, 75% vs. 20% fundus improvement and 60% vs. 30% field improvement). Conclusion: We concluded that fully endoscopic procedure result in improved rates of complete tumor removal and a reduced incidence of complications, when compared to the microscopic approach.

Endoscopic Endonasal Surgery for Clinically Nonfunctioning Pituitary Adenomas

Background: Clinically nonfunctioning pituitary adenomas (NFPAs) are the most frequent pituitary macroadenomas, and represent approximately one-third of all pituitary adenomas. Patients often present with symptoms of mass effects, such as visual field defects, chronic headache, and hypopituitarism. Objective: The objective of this study was to retrospectively analyze the surgical results of 35 patients with non-functioning pituitary adenomas (NFPAs) operated by the endoscopic endonasal approach (EEA). Surgical outcomes including postoperative complications, recurrence and the postoperative visual and endocrine profile were assessed. Results: This retrospective study included 35 patients operated for clinically nonfunctioning pituitary adenomas (NFPAs) in Al Azhar university hospitals treated by endoscopic endonasal approach (EEA) in the last 6 years. 65.7% (n = 23) were male and 34.3% (n = 12) were female. The mean age was 41.5 (range 18 - 65) years. The most common presenting symptoms were headache, with 60% of the patients having headache. Visual disturbances were the presenting symptom in 82.9% patients and endocrinological problems (panhypopituitarism) were present in 6 (17.1%) patients. The number of patients suffering from cranial nerve deficit was 2 (5.7%). Visual acuity and visual field improved in 23 (79.3%) of 29 patients. The rate of tumor recurrence/regrowth was higher in the case of giant adenomas and cavernous sinus invasion represented 11 patients (31.4%). Conclusions: Early endoscopic endonasal approach surgery of NFPAs and effective surgical decompression reduces morbidity. Visual deficit improves in two third of cases. NFPAs represent high rate of recurrence due to invasion or incomplete resection especially with giant adenomas. The recurrence rate with growth total resection (GTR) is lower than subtotal resection (STR).

Medical management of pituitary adenomas:a study of ultrastructural changes and receptor expressions

Objective The aim of this study is to analyze the ultrastructural changes and dopamine 2 receptor (D2R) expression levels of pituitary adenomas to clarify the mechanism of medical management. Methods 9 patients with prolactinomas and 4 with nonfunctioning pituitary adenomas(NFPAs) were treated with bromocriptine preoperatively. Ultrastructure changes of tumor cells were observed through electron microscopy.

The causes and prevention and treatment of cerebrospinal fluid leak during transsphenoidal pituitary adenomas surgery

Objective To analyze the causes and prevention and treatment of cerebrospinal fluid leak during transsphenoidal pituitary adenoma surgery. Methods Retrospective study of 260 cases of microscopic transsphenoidal surgery and 274 cases of endoscopic surgery for resection of the pituitary adenomas was performed. CSF leak occurred

Expressions of osteopontin and CD44v6 and their clinical significance in pituitary adenomas

Objective To explore expressions of osteopontin (OPN) and CD44v6 in pituitary adenomas and to evaluate relationship between expressions and invasiveness of tumor. Methods Immunohistochemistry was used to detect expressions of OPN and CD44v6 in 80 pituitary adenomas,and

Analysis of complications of transsphenoidal surgery for pituitary adenomas

Objective To analyze the complications of transsphenoidal operation for pituitary adenomas. Methods Postoperative complications of 323 patients underwent transphenoidal surgery for pituitary adenoma between January 2001 and December 2009 in our department were retrospectively reviewed. Results Insipidus was the most common complication after

Complications's prophylaxis in neuroendoscopic endonasal transsphenoidal surgery for pituitary adenomas

Objective To discuss the complicatiaons’s prophylaxis of removing pituitary adenomas by neuroendoscopy in tranasphenoidal surgery. Methods Clinical data of 253 cases treated by endoscopic endanasal transsphenoidal surgery were analysed. Results The total tumor removal was done in 216 (85.38%) cases,subtotal in

Study on the Expression Levels of CXCR4, CXCL12, CD44, and CD147 and Their Potential Correlation with Invasive Behaviors of Pituitary Adenomas

Objective To evaluate the factors of CXCR4, CXCL12, CD44, and CD147 as early potential diagnostic biomarkers by determining their expression levels in invasive and non-invasive pituitary adenomas. Methods Fresh pituitary adenoma specimens were collected from 35 pituitary adenoma （21 invasive and 14 non-invasive） patients who underwent surgical treatment in our Neurosurgery Department between January and April of 2009. The expression levels of CXCR4, CXCL12, CD44, and CD147 were evaluated firstly by flow cytometry, fluorescence microscopy in single cell suspensions, and then by immunohistochemical staining of paraffin tissue sections. Results Flow cytometric analyses showed that the percentage of CXCR4- and CXCL12-positive cells from invasive pituitary adenomas （IPA） was significantly higher in the single cell suspensions than that from non-invasive pituitary adenomas （nlPA） （P〈O.05）. Immunohistochemical staining revealed that CXCR4 and CXCL12 staining index scores of the invasive pituitary adenomas were significantly higher than those of the non-invasive pituitary adenomas （P〈O.05）. In contrast, neither flow cytometry nor immunohistochemical staining demonstrated significant difference between CD44 and CD147 expression levels, respectively. Conclusion Expression levels of CXCR4 and CXCL12 are correlated with the invasiveness of pituitary adenomas. Therefore, rather than CD44 and CD147, CXCR4 and CXCL12 may potentially serve as biomarkers for early detection of pituitary adenomas.

MAGNETIC RESONANCE IMAGING CHARACTERISTICS AND SURGICAL RESULTS OF ADRENOCORTICOTROPIN-SECRETING PITUITARY ADENOMAS

Objective To evaluate magnetic resonance imaging （MRI） characteristics and surgical results of adrenocorticotropin （ACTH） -secreting pituitary adenomas. Methods MRI characteristics and relationship between MRI positive rate and surgical results of 266 patients with pathologically confn＇med Cushing＇s disease were analyzed retrospectively. All patients underwent thin-section sagittal and coronal scans of the pituitary gland before and after administration of gadolinium-diethylene triamine pentaacetic acid （Gd-DTPA） on a 1. 5 Tesla MR/scanner, and dynamic enhanced MR/was performed in 39 patients. All patients underwent transsphenoidal adenomectomy. Endocrinological examinations and assessments were performed. Results Preoperative MR/revealed normal results in 41 （ 15.4% ） cases, microadenoma in 179 （67. 3% ）, macroadenoma in 42 （ 15.8% ）, and huge adenoma in 4 （ 1.5% ）. Pituitary apoplexy was found in 13 （4.9%） cases. Positive rate of ACTH-secreting adenomas was 84.6% （225/266） on MR/scans, and that of small microadenomas was 87.2% （34/39） on dynamic enhanced MRI scans. Preoperative endocrinological tests of 199 cases supported the diagnosis of typical Cushing＇s disease, while the other 67 cases had atypical endocrinological results. The endocrinological cure rate, remission rate, and inefficacy rate were 85.7%, 7.9%, and 6. 4%, respectively. There was no difference in the initial endocrinological cure rate between the patients with positive and normal MRI results （90% vs. 87. 8%, P =0. 904）. Conclusions Enhanced coronal pituitary MR/ is helpful for preoperative localization of ACTH-secreting pituitary microadenoma. Dynamic enhanced MR/may improve detection rate of microadenoma. There is no marked difference in the surgical results for patients with preoperative MR/results indicating presence or absence of microadenoma.

Overview of the 2022 WHO Classification of Pituitary Adenomas/Pituitary Neuroendocrine Tumors:Clinical Practices,Controversies,and Perspectives

The latest edition of the WHO classification of the central nervous system was published in 2021.This review summarizes the major revisions to the classification of anterior pituitary tumors.The most important revision involves preferring the terminology of pituitary neuroendocrine tumor(PitNET),even though the terminology of pituitary adenoma(PA)still can be used according to this WHO classification compared to the previous one.Moreover,immunohistochemistry(IHC)examination of pituitary-specific transcription factors(TFs),including PIT1,TPIT,SF-1,GATA2/3,and ERα,is endorsed to determine the tumor cell lineage and to facilitate the classification of PitNET/PA subgroups.However,TF-negative IHC staining indicates PitNET/PA with no distinct cell lineages,which includes unclassified plurihormonal(PH)tumors and null cell(NC)tumors in this edition.The new WHO classification of PitNET/PA has incorporated tremendous advances in the understanding of the cytogenesis and pathogenesis of pituitary tumors.However,due to the shortcomings of the technology used in the diagnosis of PitNET/PA and the limited understanding of the tumorigenesis of PitNET/PA,the application of this new classification system in practice should be further evaluated and validated.Besides providing information for deciding the follow-up plans and adjunctive treatment after surgery,this classification system offers no additional help for neurosurgeons in clinical practice,especially in determining the treatment strategies.Therefore,it is necessary for neurosurgeons to establish a comprehensive pituitary classification system for PitNET/PA that incorporates neuroimaging grading data or direct observation of invasiveness during operation or the predictor of prognosis,as well as pathological diagnosis,thereby distinguishing the invasiveness of the tumor and facilitating neurosurgeons to decide on the treatment strategies and follow-up plans as well as adjunctive treatment after surgery.

Hormone Secretion by Cell Culture of Human GH-PRL Secreting Pituitary Adenomas: Effects of Bromocriptine

Dopamine agonists effectively reduce the secretion of prolactin (PRL) in the great majority of prolactinomas and reduce the bulk of the adenomas, as well as have partial therapeutic effect on some patients with acromegaly. The inhibitory effect of bromocriptine (BC), a dopamine agonist, on growth hormone (GH) and PRL secretion of dispersed cells from the pituitary adenomas of 16 cases of acromegaly, which secret GH and PRL simultaneously, were evaluated in vitro. The significant inhibitory effects of BC on PRL secretion were found in 12 cases. It was also found that PRL secretion was strongly inhibited when GH was suppressed; on the contrary, when GH secretion was not suppressed, the production of PRL was not or weakly inhibited. The exact mechanism of the effects is nuclear so far. It is necessary to investigate, at molecular level, the etiology of GH-PRL adenomas and its response to therapeutic agents.

Expression of second mitochondria-derived activator of caspases, X-linked inhibitor of apoptosis protein, and caspase-3 in pituitary adenomas

Studies concerning correlations between pituitary adenomas and cell apoptosis have mainly focused on upstream apoptosis signaling, but seldom on downstream mediators. In the present study, second mitochondria-derived activator of caspases （Smac）, X-linked inhibitor of apoptosis protein （XIAP）, and caspase-3 protein were qualitatively analyzed using immunohistochemistry, and quantified by western blot. Smac, XIAP, and caspase-3 mRNA expressions were detected by reverse transcription-PCR. Results showed that XIAP protein and mRNA expressions were greater in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. However, Smac and caspase-3 protein and mRNA expressions were lower in the invasive pituitary adenoma group compared with the noninvasive pituitary adenoma group. In the invasive pituitary adenomas, Smac expression was positively correlated with caspase-3 protein and mRNA expression （Protein： r = 0.55, P 0.01; mRNA： r = 0.50, P 0.01）. Smac and caspase-3 expressions were negatively correlated with XIAP protein and mRNA expression （Protein： r = -0.56, -0.64, P 0.01; mRNA： r = -0.69, -0.67, P 0.01）. However, no significant differences in correlation among Smac, XIAP, and caspase-3 were detectable in noninvasive pituitary adenomas. These data indicated that high expression of XIAP and low expression of Smac and caspase-3 suppressed cell apoptosis and led to enhanced invasiveness of pituitary adenomas. Thus, Smac, XIAP, and caspase-3 may be useful markers in determining the invasive behavior of pituitary adenomas.

The Expression of Integrinβ1 and FAK in Pituitary Adenomas and Their Correlation with Invasiveness

The expression and possible role of integrin-focal adhesion kinase signal pathway in invasive pituitary adenomas were explored. Forty-nine human pituitary adenomas were detected for the expression of integrinβ1 （INTβ1） and focal adhesion kinase （FAK） by immunohistochemistry, and their correlation with the invasiveness of pituitary adenomas as well as between themselves was ana- lyzed. The results showed that INTβ1 was expressed in 46 cases （93.9%） and FAK in 36 cases （73.5%）, respectively, and their expression levels were highly correlated with tumor invasiveness, but not with the tumor types. It was suggested that the integrin-focal adhesion kinase signal pathway plays a role in the invasiveness of pituitary adenomas.

IN SITU PCR AND IMMUNOHISTOCHEMICAL STUDIES ON p16 GENE IN PITUITARY ADENOMAS

Objective: To examine the occurrence of p16 gene deletion and to analyze p16 expression on paraffin-embedded human pituitary adenoma specimens. Efforts were made to optimize the technical conditions forin situ PCR. Methods:In situ PCR techniques and inimuno-histochemistry were used. Results: Immunohistochemically, p16-positive tumor cells were observed in all cases with various proportions. The majority of the stromal cells and part of tumor cells was devoid of p16 immunostaining, but signal ofin situ PCR for p16 gene, exon 2, was displayed in these cells. Conclusion: The results implied that p16 gene might not be deleted in these pituitary adenomas. It also indicated thatin situ PCR, both direct and indirect methods, proved feasible and informative to the aim of DNA detection. It is critical to overcome non-specific amplification in directin situ PCR by means of higher annealing temperature, fewer cycle, lower magnesium concentration and stringent washing. A target DNA-deleted sample as the negative control is extremely necessary. For the indirect method, the way to improve the sensitivity is to loosen the conditions for amplification and washing, so that more amplification products are subject to hybridization, and signal detection is facilitated.

Treatment and Prognosis of Pituitary Adenomas in Children

In order to explore the treatments and prognosis of pituitary adenomas in children, the experience in the treatment of 15 children with pituitary adenomas was summed up. The clinical data of 15 children with pituitary adenomas in Tongji Hospital were retrospectively analyzed from Jan. 1997 to Aug. 2004, including 9 males and 6 females with the age ranging from 9-15 years old （mean 12.8）. The disease duration was 3 months to 5 years （mean 2 years）. Fourteen cases were subjected to the microsurgical operation through oral tanssphenoidal approach and one case to medical treatment. All cases were followed up from 1 to 5 years by imaging studies and endocrine investigation. The results showed that 13 eases obtained total removal of the tumor and one subtotal removal of tumor. No severe complications and operation death occurred. It was concluded that the tanssphenoidal approach was the optimal treatment for children with pituitary adenomas. Regular and long-term follow-up is of great importance.