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PubMed2026

A Comparative Analysis of the Methylation Status of Non-Coding RNA Promoters in Fibroid and Matched Myometrium.

Uterine fibroids exhibit dysregulated expression of non-coding RNAs (ncRNAs), although the underlying mechanisms remain incompletely understood. We investigated promoter DNA methylation and its relationship with ncRNA expression in fibroids. Genomic DNA from eight paired fibroid and matched myometrial tissues was analyzed using MeDIP-chip to identify differentially methylated ncRNA promoters. Selected candidates were validated by methylation-specific PCR (MSP) in 16 paired samples, and transcript expression was assessed by qRT-PCR in 68-94 paired specimens. MeDIP-chip identified 538 lncRNAs and 61 miRNAs with differential promoter methylation, including 300 hypermethylated and 238 hypomethylated lncRNAs and 47 hypermethylated and 14 hypomethylated miRNAs. Promoter methylation was not significantly correlated with transcript expression (r = -0.1224). MSP confirmed hypermethylation of LINC-PINT and MIR9-3 and hypomethylation of WT1-AS and TTLL10-AS1. Correspondingly, LINC-PINT and MIR9-3 expression was decreased, whereas WT1-AS and TTLL10-AS1 expression was increased in fibroids. However, LINC-PINT and TTLL10-AS1 methylation did not fully correspond with MeDIP-chip findings. These results reveal widespread ncRNA promoter methylation alterations in uterine fibroids but demonstrate that genome-wide methylation does not consistently predict transcript expression, highlighting the complexity of ncRNA epigenetic regulation and the importance of locus-specific validation.

Humans↗
PubMed2025

Genome-Wide Analysis of the AT-Hook Gene Family in Malus sieversii and Functional Characterization of MsAHL13.

AT-hook motif nuclear-localized (AHL) proteins are pivotal in plant growth, development, and stress responses. Nevertheless, there is limited research on AHL proteins in Malus sieversii. Our study identified 25 AHL genes from the M. sieversii genome, named MsAHL1-MsAHL25. The encoded protein sequences had lengths ranging from 195 to 554 amino acids, molecular weights from 19.17 to 58.53 kDa, and isoelectric points from 4.67 to 10.09. Chromosomal mapping revealed that these 25 genes were unevenly distributed across 10 chromosomes. Collinearity analysis of AHL genes in M. sieversii implied that gene loss might have occurred during its evolution. The phylogenetic tree classified the AHL proteins of M. sieversii into two subfamilies, showing a close relationship with multiple proteins of M. domestica. Promoter analysis indicated that the AHL genes in M. sieversii harbored numerous stress- and hormone-responsive elements, suggesting their potential role in various stress responses. qRT-PCR analysis of six representative MsAHLs under biotic and abiotic stresses demonstrated that the expression of MsAHL13, MsAHL15, and MsAHL17 was significantly upregulated under salt, drought, and cold stresses, while MsAHL01 expression was inhibited under low-temperature stress. All six MsAHLs were induced by the pathogen Valsa mali. Subcellular localization analysis of the specifically expressed protein MsAHL13 showed its nuclear location. Furthermore, luciferase and yeast two-hybrid assays confirmed the in vitro physical interaction between the MsAHL13 and MsMYB1 proteins. This research offers an important theoretical basis for further exploration of the functional mechanisms of this gene family in responding to environmental stresses.

Malus sieversii↗
PubMed2026

Genome-Wide Characterization and Salt-Responsive Expression Divergence of Chromosome Group 2 and Group 6 TaBADH Genes in Wheat.

Betaine aldehyde dehydrogenase (BADH) catalyzes the final step in glycine betaine biosynthesis, but the evolutionary divergence and differential salt responsiveness of BADH homeologs in bread wheat remain unclear. We identified six TaBADH genes and analyzed their phylogenetic relationships, conserved motifs, gene structures, promoter cis-acting elements and synteny. RNA-seq and qRT-PCR were used to compare expression in salt-tolerant Jimai 60 and salt-sensitive Chinese Spring under 200 mM NaCl, and BADH activity, glycine betaine, H2O2 and malondialdehyde (MDA) were measured during treatment. The genes separated into chromosome group 2 and group 6 clades with distinct structural and transcriptional patterns. TaBADH-2B encoded a shorter protein and lacked several conserved motifs. Group 6 genes showed stronger salt-responsive expression in Jimai 60, with TaBADH-6D displaying the strongest and most sustained induction. Jimai 60 also showed higher BADH activity and glycine betaine accumulation and lower H2O2 and MDA contents at later time points. Expression of TaBADH-6D improved E. coli growth under 200 mM NaCl. These findings identify homeolog-specific divergence within the BADH wheat family and support TaBADH-6D as a candidate for plant-level functional validation.

TaBADH-6D↗
PubMed2026

Genome-Wide Characterization of the Apple HD-Zip IV Gene Family and Functional Validation of MdHDZIV3 Under PEG-Induced Osmotic Stress.

The homeodomain-leucine zipper IV (HD-Zip IV) transcription factor subfamily plays essential roles in epidermal development, cuticle formation, lipid metabolism, and environmental adaptation in plants. Despite its biological importance, the HD-Zip IV family has not been systematically characterized in apple (Malus domestica). Here, we identified 17 apple HD-Zip IV genes and named them MdHDZIV1-MdHDZIV17 based on their locations on the chromosomes. The 17 genes showed a nonuniform distribution on eight chromosomes, while the occurrence of both tandem and segmental duplications indicated that family expansion involved more than one duplication mechanism. All MdHDZIV proteins contained the conserved HD, LZ, START, and SAD domains but lacked the MEKHLA domain, consistent with typical HD-Zip IV structural features. Phylogenetic analysis classified MdHDZIV proteins into five groups together with HD-Zip IV members from Arabidopsis thaliana and rice, indicating evolutionary conservation of this subfamily. Collinearity and Ka/Ks analyses revealed that duplicated MdHDZIV gene pairs were mainly subjected to purifying selection. Promoter scanning revealed diverse cis-regulatory motifs associated with hormonal signaling, environmental stress, light response, and epidermal regulation, including ABRE, ARE, W-box, MYC, G-box, and L1-box motifs. Integration of transcriptomic profiling with qRT-PCR validation revealed pronounced tissue-dependent differences in the expression of MdHDZIV genes in leaf, fruit skin, and branch bark. Under PEG6000-induced osmotic stress and NaCl-induced salt stress, 10 candidate MdHDZIV genes displayed gene-specific and stress type-specific expression patterns, with MdHDZIV3 showing strong induction under PEG6000 treatment. Functional validation in apple calli showed that MdHDZIV3 overexpression enhanced PEG tolerance, increased fresh weight, elevated SOD and POD activities, and reduced MDA accumulation under osmotic stress. These findings provide a genome-wide framework for understanding the apple HD-Zip IV gene family.

abiotic stress↗
PubMed2026

Common Molecular Mechanisms and Candidate Drug Targets in Type 2 Diabetes Mellitus and Atherosclerotic Cardiovascular Disease.

This study examined the mechanisms underlying the comorbidity between type 2 diabetes mellitus (T2DM) and atherosclerotic cardiovascular disease (ASCVD), while identifying potential therapeutic targets. Common differentially expressed genes (C-DEGs) between T2DM and ASCVD were extracted from the GSE78721 and GSE12288 datasets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses, protein-protein interaction (PPI) network construction, hub gene identification, and Drug-Gene Interaction Database (DGIdb) analysis were conducted. The association between hub C-DEGs and immune-infiltrating cells was analyzed using the CIBERSORT method. Expression levels of hub C-DEGs were quantified through qRT-PCR and Western blot analyses. A total of 32 C-DEGs were identified, comprising 20 upregulated and 12 downregulated genes. C-DEGs were predominantly enriched in key pathways, including viral myocarditis, arrhythmogenic right ventricular cardiomyopathy, hypertrophic cardiomyopathy, and dilated cardiomyopathy. PPI analysis revealed 29 nodes and 39 edges, leading to the identification of eight hub C-DEGs (HSP90B1, PLAU, SLPI, TOP3A, NCF4, PRF1, TUBA1C, and CS) across both datasets. Furthermore, hub C-DEGs (TOP3A, SLPI, NCF4, PRF1, and PLAU) demonstrated significant correlations with immune-infiltrating cell levels. Drugs specifically targeting these hub C-DEGs present promising candidates for the treatment of T2DM and ASCVD. Additionally, the expression of hub C-DEGs at both mRNA and protein levels was validated in patients with T2DM and ASCVD. An integrated bioinformatics analysis facilitated the screening of candidate therapeutic targets, mechanisms, and drugs for T2DM and ASCVD, offering new insights into molecular therapies for these conditions.

Diabetes Mellitus, Type 2↗
PubMed2026

Proteomic insights into azoospermia: protein differences in testicular tissue between non-obstructive and obstructive azoospermia patients.

Non-obstructive azoospermia (NOA) and obstructive azoospermia (OA) are the main classifications of severe male infertility, but the molecular mechanism of NOA remains poorly understood. This study aimed to identify potential biomarkers and pathological mechanisms by comparing the proteomic differences in testicular tissues of NOA and OA patients. Through proteomic analysis based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) of testicular samples from 5 NOA patients and 5 OA patients, we identified 5264 proteins, among which 717 differentially expressed proteins (DEPs) were found between the two groups (242 upregulated and 475 downregulated in NOA). Bioinformatics analysis indicated that these DEPs were significantly associated with reproductive development, gametogenesis, and cell structural stability. On the basis of this, six candidate proteins, including dysferlin (DYSF), myoferlin (MYOF), mitsugumin 53 (MG53), cluster of differentiation 63 (CD63), caveolin-3 (CAV3), and calpain-3 (CAPN3), were selected from the DEPs and verified in an expanded sample set (37 NOA cases and 28 OA cases) through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, confirming their dysregulation in NOA. These findings provide new proteomic insights into NOA, highlighting the disruption of membrane repair and structural pathways, and offer potential biomarkers for understanding its pathogenesis.

Humans↗
PubMed2000

[Profiles of expression of genes coding kininogen and kinin receptors as a marker of tissue pathology in cervical cancer coexisting with HPV infection].

Kinins are peptides involved in inflammatory processes, vascular permeability, proliferation and mitogenesis of tumor cells. The majority of kinins actions are mediated through an interaction with cell surface bradykinin receptors BR1 and BR2. Kinins precursor is kininogen (kng). The changes in proteins are initiated by changes in the expression of genes coding these proteins, thus can be a valuable diagnostic markers of malignant processes including cervical carcinoma. The paper presents an analysis of kininogen-kinins receptor genes expression in women treated surgically because of a carcinoma of uterine cervix. Among the studied women in 5 cases previously brachyHDR therapy was applied. In all studied cases HPV 18 infection and in 2 cases a co-infection HPV 16/18 by use of Consensus Primers MY09, MY 11 and type specific primers for HPV 16, 18 was ascertained. In RNA extracts the number of the mRNA copies for kiningen, BR1 and BR2 was assessed using QRT-PCR Taq Man. The higher expression of BR1 than BR2 was marked in the tissue with cancer cells. In the patients after brachytherapy higher expression of BR2 than BR1 mRNA was found. The higher BR1 expression was also shown in iliac lymph nodes in patients with active neoplastic process, opposite to the patients after brachytherapy in whom higher BR2 expression was ascertained. The lack of expression of kng mRNA was found only in 3 specimens. The high expression of kinin receptors especially BR1 in infiltrating carcinoma margin can be a marker of pathology intensity: proliferative potential of neoplasms cells or chronic inflammatory state in the presence of invasive carcinoma.

Adult↗
PubMed2001

Quantitative RT-PCR assay for mRNA of VEGF and histone H4 in the determination of proliferative and angiogenic activity in vulvar pathology.

Proliferative and angiogenic activity of tissue specimens taken from women with various vulvar pathologies were evaluated by determining the number of mRNA VEGF molecules and H4 histone mRNA molecules, by means of the QRT-PCR (TaqMan) technique. Following a cluster analysis the results, where normalised. Euclidean distances were used, all the cases were classified into three groups of pathologies. Group I included low degree vulvar pathologies, group II included high degree vulvar pathologies and group III included vulvar pathologies with high proliferative and angiogenic activity. Significant differences were found in the proliferative and angiogenic activity between groups I and III, and between groups II and III, while no statistically significant differences were found between groups I and II.

Carcinoma in Situ↗
PubMed2001

Differentiation strategies of alternative splicing variants mRNA estrogen receptor-alpha.

From the clinical point of view recognizing the concentrations and type profile of isoforms could be of significant practical importance. The aim of the study is designed QRT-PCR reaction to assess profile of mRNA ER-alpha and their isoforms. Theoretical part of the study was made according to computer program and available Genbank database. To detect a isoform one of the primer was designed to hybridize within exon-border linked in alternative splicing. The study presents the differentiation strategies in isoforms coming about as the alternative splicing result. Designed oligonucleotide probes and primers allow to distinguish mRNA isoforms of ER-alpha and their quantification in assessed tissues.

Alternative Splicing↗
PubMed2001

Quantitative analysis of estrogen receptor-alpha and -beta and exon 5 splicing variant mRNA in endometrial hyperplasia in perimenopausal women.

We used real-time reverse transcription-polymerase chain reaction (QRT-PCR) to detect wild-type ERalpha and ERbeta and their isoforms as well as exon 5 splicing variant mRNA ERalpha in various types of endometrial hyperplasia in perimenopausal women. Our data suggest that the exon 2 and 7 deletion ERalpha mRNA isoforms are present in endometrial tissue undergoing various hyperplastic states. Wild type ERbeta mRNA and their isoforms 2, 4, 5, 6 were observed in each type of assessed endometrial specimens, and ERalpha/delta5 was found only in hyperplasia complex group.

Adult↗
PubMed2002

[Time-dependent effects of interleukin-8 gene expression in endothelial cells exposed on fluid shear stress].

Fluid shear stress plays an important role in vascular biology. In vivo, endothelial cells are continuously exposed to mechanical shear stress generated by the flowing blood. Previous studies have identified the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expression of many genes involved in vascular physiology and pathophysiology. To investigate the role of fluid shear stress on IL-8 expression in human umbilical vein endothelial cells (HUVECs), we employed quantitative reversal transcription-polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. Here we show that IL-8 mRNA did not express in HUVECs untreated with fluid shear stress. IL-8 mRNA expression increased when HUVECs exposed to fluid shear stress for 1 h, and it reached the summit when HUVECs exposed to fluid shear stress for 2 h. Then IL-8 expression gradually decreased at 3 h of stimulation by shear stress and remained at a constant level throughout the time course of the study. The increase of IL-8 expression by shear stress was time-dependent. The biphasic response of IL-8 gene expression was found in experiments in which the applied shear stress was 2.23 dyne/cm2, 4.20 dyne/cm2, or 6.08 dyne/cm2. IL-8 gene expression in response to shear stress was very similar to NF-kappa B in response to shear stress. The induction of IL-8 gene expression by fluid shear stress is probably due to the activation of NF-kappa B. This in vitro study demonstrates the expression of IL-8 gene can be regulated by shear stress. Fluid shear stress induces a biphasic response of human IL-8 gene expression in HUVECs. These considerations suggest that IL-8 expression induced by fluid shear stress in HUVECs may play an important role in the genesis and development of both inflammation and arterioatherosclerosis.

Cells, Cultured↗
PubMed2002

[Force-dependent effects of interleukin-8 gene expression in endothelial cells exposed on fluid shear stress].

Fluid shear stress plays an important role in many physiological and pathophysiological processes of the cardiovascular system. Previous studies have identified the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expression of many genes, including IL-8 gene expression, and IL-8 expression induced by fluid shear stress was time-dependent. To investigate the role of intensity of fluid shear stress on IL-8 expression in human umbilical vein endothelial cells (HUVECs), we had HUVECs exposed to shear stress 2.23, 4.20, 6.08, 8.19, 9.67, 12.15, 14.40, 16.87, and 19.29 dyne/cm2 respectively and employed quantitative reversal transcription-polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. The results show that IL-8 mRNA did not express in HUVECs untreated with fluid shear stress. The IL-8 mRNA expression by shear stress was force intensity-dependent. After HUVECs exposed to low fluid shear stress (2.23 dyne/cm2) for 1 h or 2 h, IL-8 mRNA expression increased near 68 or 52 times as that of HUVECs exposed to high fluid shear stress (19.29 dyne/cm2). The linear regression equations between IL-8 mRNA expression (log (copies), y) and shear stress (dyne/cm2, x) are: y = 7.57 - 0.11x, r = 0.97 (for 1 h); y = 7.92 - 0.10x, r = 0.96 (for 2 h). This in vitro study demonstrates the expression of IL-8 gene can be regulated by fluid shear stress. The low shear stress could induces much more expression of IL-8 mRNA, which plays probably an important role in the pathogenesis of inflammation and arteroatherosclerosis.

Cells, Cultured↗
PubMedDate not supplied

Induction of Epstein-Barr virus (EBV) reactivation in Raji cells by doxorubicin and cisplatin.

BACKGROUND: Epstein-Barr virus (EBV) can be activated in B-lymphoid cells to enter the lytic cycle by various kinds of stimuli, including 12-O-tetradecanoylphorbol-1 3-acetate (TPA), butyric acid, calcium ionophore A23187, transforming growth factor-beta and anti-immunoglobulin crosslinking. EBV reactivation has been clinically observed in patients receiving systemic chemotherapy. This study sought in vitro evidence to suggest whether anticancer drugs may directly contribute to the EBV reactivation. MATERIALS AND METHODS: Raji cells, an EBV-containing Burkitt's lymphoma cell line, were used as the experimental model. TPA served as a positive control for chemical induction of EBV reactivation. Expression of the BZLF1 transcript of EBV and its encoded protein, ZEBRA, were examined by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and flow cytometry, respectively. Transactivation activity of ZEBRA was further assessed by a luciferase reporter assay of EBV DR-promoter activity and a flow cytometry assay assessing the endogenous expression of EA-D (BMRFl). RESULTS: Doxorubicin and cisplatin, two commonly used anticancer agents, induced a dose-dependent up-regulation of BZLF1 mRNA and ZEBRA protein. The luciferase reporter activity and the expression of endogenous EA-D protein, also increased by doxorubicin and cisplatin, indicated an up-regulation of the transactivating activity of ZEBRA. CONCLUSION: These data indicate that cytotoxic anticancer drugs may up-regulate the expression and the transactivating activity of BZLF1, and suggest that systemic chemotherapy may be a risk factor for EBV reactivation in patients with EBV-associated malignancies.

Antigens, Viral↗
PubMed2003

[Expression patterns in isoforms of vascular endothelial growth-factors in tissue samples of vulval cancer T1 N2M0 stage].

OBJECTIVES: The aim of the study was estimating the expression of isoforms of mRNA vascular endothelial growth factor: VEGF121, VEGF145, VEGF165, VEGF183, VEGF189, VEGF206, in tissue samples of vulvar cancer, normal and inguinal lymph nodes metastases. MATERIALS AND METHODS: The material for this investigation of expression of mRNA selected genes was 4 tissue specimens from women with vulvar cancer T1N2M0. The method used for quantitative determination of the numbers of mRNA copies of selected genes was the QRT-PCR and QPCR technique, with the use of an ABI PRISM 7700 (TaqMan) sequence detector. The nucleotide sequence for starters and probes for quantitative RT-PCR was designed with the use of the Primer Express Version 1.0 ABI PRISM software. RESULTS: Expression of all mRNA isoforms VEGF was found in the samples. The number of mRNA copies obtained in 1 microgram total mRNA extract of the genes examined in tissue sections with various pathomorphology were clearly differentiated. CONCLUSIONS: Expression profile of aforementioned genes indicated a definitive relationship between vascular endothelial growth factor isoform and vulvar cancer. These findings provide evidence that the expression pattern of mRNA VEGF isoform should find broad applications in clinical and research settings.

Endothelial Growth Factors↗
PubMed2002

Transcriptional analysis of tumor-specific T-cell responses in cancer patients.

Over the last decade, tumor immunology in general and tumor immunotherapy in particular have made important progress. Thanks to the discovery of tumor-associated antigens (TAA), and the consequent development of active-specific immunization protocols, TAA-specific T-cell responses can now be induced reproducibly in cancer patients. However, clinical responses directly ascribable to TAA-specific T cells occur only occasionally. It is not clear why TAA-specific T cells do not eliminate tumor cells in vivo. This paradoxical coexistence of antigen-bearing tumor cells and antigen-specific T cells constitutes a critical point of current investigation. In recent years, protein-ligand interaction-based methods aimed at the identification and characterization of T cells using antibodies (cell surface phenotyping, intracellular and secreted cytokine detection), or HLA-peptide multimers, have significantly improved the analysis of TAA-specific T cells. These methods, however, seem to have reached the limit of their usefulness. Transcriptional analysis may add sensitivity and resolution and may provide global pictures of the multifactorial requirements for an efficient immune response against tumors. In this review, we describe the use of molecular genetic methods, such as real time qRT-PCR, cDNA microarrays, and TCR repertoire analysis. In addition, we describe techniques for high-fidelity messenger RNA amplification that allow high-throughput analysis of samples obtained from minimal sources, such as HLA/peptide tetramer sorted antigen-specific T cells, laser capture dissection, or fine needle aspirates. Recent work discussed in this review summarizes the complementarity of transcriptional analysis as an essential tool that, in addition to conventional methods, may deepen and broaden the characterization of tumor-specific T cells.

Gene Expression Profiling↗
PubMed2003

Selectively reduced expression of synaptic plasticity-related genes in amyloid precursor protein + presenilin-1 transgenic mice.

A critical question in Alzheimer's disease (AD) research is the cause of memory loss that leads to dementia. The amyloid precursor protein + presenilin-1 (APP+PS1) transgenic mouse is a model for amyloid deposition, and like AD, the mice develop memory deficits as amyloid deposits accumulate. We profiled gene expression in these transgenic mice by microarray and quantitative RT-PCR (qRT-PCR). At the age when these animals developed cognitive dysfunction, they had reduced mRNA expression of several genes essential for long-term potentiation and memory formation (Arc, Zif268, NR2B, GluR1, Homer-1a, Nur77/TR3). These changes appeared to be related to amyloid deposition, because mRNA expression was unchanged in the regions that did not accumulate amyloid. Transgene expression was similar in both amyloid-containing and amyloid-free regions of the brain. Interestingly, these changes occurred without apparent changes in synaptic structure, because a number of presynaptic marker mRNAs (growth-associated protein-43, synapsin, synaptophysin, synaptopodin, synaptotagmin, syntaxin) remained stable. Additionally, a number of genes related to inflammation were elevated in transgenic mice, primarily in the regions containing amyloid. In AD cortical tissue, the same memory-associated genes were downregulated. However, all synaptic and neuronal transcripts were reduced, implying that the loss of neurons and synapses contributed to these changes. We conclude that reduced expression of selected genes associated with memory consolidation are linked to memory loss in both circumstances. This suggests that the memory loss in APP+PS1 transgenic mice may model the early memory dysfunction in AD before the degeneration of synapses and neurons.

Aged↗
PubMed2003

[Differential expression of the epithelial membrane protein 1 of laryngeal carcinoma].

OBJECTIVE: In order to explore pathogenic mechanism of laryngeal carcinoma, the involved genes were identified in larynx carcinogenesis by comparing the gene expression profile in matched primary normal epithelial cells and primary laryngeal carcinoma cells from the same patients. METHODS: A cDNA microarray analysis consisting of 11,431 human genes revealed significant changes in the expression of 35 genes, with 8 genes being up-regulated and 27 being down-regulated. The epithelial membrane protein-1 (EMP-1) is one of the down-regulated genes. EMP-1 expression in various kinds of laryngeal carcinoma was determined by semi-quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). RESULTS: The EMP-1 mRNA levels in all laryngeal carcinoma cells was significantly lower than that in the matched primary normal epithelial cells (P < 0.05) and were not correlated to the stage and differentiation of laryngeal carcinoma (P > 0.05). CONCLUSIONS: The EMP-1 expression was correlated to larynx carcinogenesis and may be helpful to elucidate the pathogenic mechanism in laryngeal carcinoma.

Adult↗
PubMed2003

[Evaluation of wild-type estrogen receptor beta and its isoforms: ER-beta/delta 5/6 i ER-beta/delta 6 in endometrial adenocarcinoma].

Estrogen target genes play essential role among pathogenetic and prognostic factors of endometrial adenocarcinoma; and among them estrogen receptor genes. Although prevailing estrogen receptor type is ER-alpha in endometrium, transduction of signal carried by estrogens can be also mediate by ER-beta, which biological meaning relies on modulation of ER-alpha transcriptional activity. There is hypothesized that ER-beta can keep under control mitogenic activity mediating by ER-alpha. The aim of the study was qualitative and quantitative analysis of wtER-beta and ER-beta/delta 5-6, ER-beta/delta 6 isoforms in endometrial adenocarcinoma. Material used in the study included specimens obtained from 27 female: group I--normal endometrium (n = 12), group II--endometrial adenocarcinoma (n = 15). RNA was extracted from the analyzed material using phenol-chloroform method by Total RNA Prep Plus. Qualitative analysis was performed basing on RT-PCR reaction and polyacrylamide gel electrophoreses. For all RNA extracts of the samples examined RT-PCR was performed with a sequence detector ABIPRISMTM7700--Perkin-Elmer Applied Biosystems (RT: 600 C-30 min; PCR: 950 C-5 min, 40 cycles: 950 C-30s, 600 C-1 min, and 720 C-10 min) in order to determine the number of RNA copies (which corresponds to the number of cDNA copies) for wtER-beta and ER-beta/delta 5-6 and ER-beta/delta 6. Specificity of QRT-PCR reaction was determined by delimitation of melting temperature of all examined amplimers (ABI PRISM7000, Qu-antiTect SYBR Green RT-PCR Kit) and by sequence analysis using ABI PRISM377 DNA Sequencer. Wild-type of estrogen receptor beta and ER-beta/delta 5-6, ER-beta/delta 6 isoforms--coming into alternative splicing of mRNA, presented both, in proliferative endometrium and endometrial adenocarcinoma. Copy number of wtER-beta mRNA was significantly (p < 0.05, Mann-Whitney U test) lower in comparison with proliferative endometrium and correlated with decrease of ER-beta/delta 6 mRNA copy number (r = 0.85; p < 0.05). Preliminary results confirming decrease of wtER-beta, ER-beta/delta 5/6 and ER-beta/delta 6 mRNA copy number in endometrial adenocarcinoma can show their relationship with high risk of carcinogenesis.

Adenocarcinoma↗