scholarly journals Aberrant Expression of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) in Warthin Tumors

Cancers ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1112 ◽  
Author(s):  
Robert Mandic ◽  
Abbas Agaimy ◽  
Daniel Pinto-Quintero ◽  
Katrin Roth ◽  
Afshin Teymoortash ◽  
...  
Keyword(s):  
Parotid Gland ◽  
Normal Control ◽  
Staining Pattern ◽  
Rna Extraction ◽  
Renal Oncocytoma ◽  
Mrna Levels ◽  
Warthin Tumor ◽  
Aberrant Expression ◽  
Ductal Cells ◽  
Gapdh Mrna

The Warthin tumor represents the second most frequent benign tumor of the parotid gland and is characterized by the presence of oncocytes rich in structurally and functionally altered mitochondria. Next to its role in metabolism, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is also implicated in cellular mitophagy. Immunohistochemistry was carried out on Warthin tumor and normal control (parotid gland with striated ducts) tissues, using anti-GAPDH specific antibodies followed by digital image analysis. Laser capture microdissection was used to isolate the oncocytic tumor cell and normal control striated duct compartments for RNA extraction and qPCR. Warthin tumor oncocytes exhibited a markedly spotted GAPDH staining pattern exhibiting cells with cytoplasmic and nuclear, only nuclear or none GAPDH staining. A significantly lower (p < 0.0001) total GAPDH signal was detected in Warthin tumor oncocytes. Similarly, significantly lower (p < 0.005) GAPDH mRNA levels were seen in oncocytes compared with normal ductal cells. To exclude the possibility of this GAPDH staining pattern being a general feature of oncocytic neoplasms of different organs, we tested a cohort of renal oncocytoma and oncocytic chromophobe carcinoma; none showed this type of staining. The observed progressive GAPDH loss in Warthin tumor oncocytes could be implicated in the pathogenesis of Warthin tumors.

scholarly journals NPY and sbGnRH gene expression in juvenile and adult male Brazilian flounder Paralichthys orbignyanus

2011 ◽  
Vol 41 (11) ◽  
pp. 1927-1930 ◽  
Author(s):  
Vinicius Farias Campos ◽  
Tiago Veiras Collares ◽  
Fabiana Kömmling Seixas ◽  
João Carlos Deschamps ◽  
Luis Fernando Fernandes Marins ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Extraction ◽  
Sea Bream ◽  
Mrna Levels ◽  
Adult Males ◽  
Adult Fish ◽  
Cdna Synthesis ◽  
Hypothalamic Regulation ◽  
Measure Gene Expression ◽  
Paralichthys Orbignyanus

The objective of this study was to evaluate neuropeptide Y (NPY) and sea bream gonadotropin-release hormone (sbGnRH) gene expression in juvenile and adult males of Brazilian flounder. Hypothalamuses from fish were sampled for total RNA extraction. After cDNA synthesis, real-time PCR was used to measure gene expression. NPY showed approximately 2-fold increases in their mRNA levels while sbGnRH showed 3-fold increases in adult fish. These results suggest that these peptides could be involved on hypothalamic regulation of Brazilian flounder sexual maturation.

Stability of housekeeping genes and expression of locally produced growth factors and hormone receptors in goat preantral follicles

Zygote ◽  
2010 ◽  
Vol 19 (1) ◽  
pp. 71-83 ◽  
Author(s):  
Isana M. A. Frota ◽  
Cintia C. F. Leitão ◽  
José J. N. Costa ◽  
Ivina R. Brito ◽  
Robert van den Hurk ◽  
...  
Keyword(s):  
Growth Factors ◽  
Hormone Receptors ◽  
Rna Extraction ◽  
Housekeeping Genes ◽  
Stable Gene ◽  
Mrna Levels ◽  
Preantral Follicles ◽  
The Stability ◽  
Suitable Reference

SummaryThe aim of the present study was to investigate the stability of six housekeeping genes, and the relative expression of growth factors (EGF, GDF-9, BMP-15, VEGF, FGF-2, BMP-6, IGF-1 and KL) and hormone receptors (FSH, LH and GH) in goat preantral follicles. To evaluate to stability of housekeeping genes micro-dissected fresh follicles (150–200 μm) as well as follicles that have been in vitro cultured for 12 days were used. In addition, isolated fresh follicles were used to compare expression of various growth factors and hormone receptors before culture. Both fresh and cultured follicles were subjected to total RNA extraction and synthesis of cDNA. After amplification of cDNA by real-time PCR, the geNorm software program was used to evaluate the stability of glyceraldehyde-2-phosphate dehydrogenase (GAPDH), β-tubulin, β-actin, phosphoglycerokinase (PGK), 18S rRNA, ubiquitin (UBQ) and ribosomal protein 19 (RPL-19). In addition, follicular steady-state levels of mRNA from the various growth factors under study were compared. Results demonstrated that, in goat preantral follicles, UBQ and β-actin were the most suitable reference genes and thus could be used as parameters to normalize data from future in vitro studies. In contrast, 18S RNA appeared the least stable gene among the tested housekeeping genes. Analysis of mRNA for several hypophyseal hormone receptors in fresh preantral follicles showed significantly higher FSH-R mRNA levels than those of LH-R and GH-R, and no difference between GH-R and LH-R mRNA levels. In regard growth factor mRNA expression in goat preantral follicles, EGF mRNA levels appeared significantly lower than those of the other studied growth factors. Increasingly higher relative mRNA levels were observed for GDF-9, BMP-15, BMP-6, FGF-2, VEGF, Kl and IGF-1, successively. In conclusion, UBQ and β-actin are the most stable housekeeping genes in fresh and 12-days cultured caprine preantral follicles. Furthermore, in fresh follicles, high levels of FSH-R mRNA are detected while among eight growth factors, IGF-1 is the most highly expressed and EGF the weakest expressed compound.

Studies of Muc-1 mucin expression and polarity in the mouse mammary gland demonstrate developmental regulation of Muc-1 glycosylation and establish the hormonal basis for mRNA expression

1992 ◽  
Vol 101 (1) ◽  
pp. 191-199 ◽  
Author(s):  
G. Parry ◽  
J. Li ◽  
J. Stubbs ◽  
M.J. Bissell ◽  
C. Schmidhauser ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cdna Probe ◽  
Mouse Mammary Gland ◽  
Mammary Development ◽  
Mammary Epithelial ◽  
Mrna Levels ◽  
Apical Surface ◽  
Ductal Cells ◽  
Carbohydrate Epitopes ◽  
Mucin Expression

Muc-1 is a major mucin glycoprotein expressed on the surface of mammary epithelial cells. It has attracted considerable attention as it is expressed in an aberrant form on many breast tumor cells. Here we describe studies using a recently obtained cDNA probe of Muc-1 expression during lactogenic development in the mouse. Northern blot analysis demonstrated that Muc-1 is expressed at all stages of lactogenic development but its levels are increased very significantly during mid-pregnancy and into lactation. The basis of this was examined using CID-9 mammary epithelial cell cultures. It was found that in the presence of insulin Muc-1 mRNA levels were increased by both hydrocortisone and prolactin, with the combination of the three hormones supporting maximum expression. Muc-1 mRNA levels were also modulated by culturing cells on a basement-membrane-like extracellular matrix that promoted mRNA levels 5- to 10-fold above levels in cells cultured on plastic tissue culture dishes. Immunocytochemical studies using monoclonal antibodies to carbohydrate epitopes on Muc-1 demonstrated that while Muc-1 was found at all developmental stages, it became increasingly sialylated during the course of pregnancy and into lactation. Additionally, we found that while Muc-1 is tightly polarized to the apical surface of the epithelium of lactating and pregnant mice it exhibited a less-polarized distribution on a small proportion of ductal cells in virgin mice. We conclude that the expression of Muc-1 is regulated at several different levels and by a number of different factors. We speculate that this may reflect different functional roles for Muc-1 at different stages of mammary development.

scholarly journals Basal cell adenoma and myoepithelioma of the parotid gland: patterns of enhancement at two-phase CT in comparison with Warthin tumor

2019 ◽  
Vol 25 (4) ◽  
pp. 285-290 ◽  
Author(s):  
Ji Young Lee ◽  
◽  
Hyung-Jin Kim ◽  
Yi Kyung Kim ◽  
Jihoon Cha ◽  
...  
Keyword(s):  
Parotid Gland ◽  
Basal Cell ◽  
Basal Cell Adenoma ◽  
Warthin Tumor ◽  
Two Phase ◽  
Cell Adenoma

Effect of neoadjuvant chemoradiotherapy on p53 and SOX2 protein expression in esophageal adenocarcinoma

2020 ◽  
Vol 14 (9) ◽  
pp. 785-793
Author(s):  
Lucia Suzuki ◽  
Daan Nieboer ◽  
Jan JB van Lanschot ◽  
Manon CW Spaander ◽  
Leendert HJ Looijenga ◽  
...  
Keyword(s):  
Protein Expression ◽  
Esophageal Adenocarcinoma ◽  
Staining Pattern ◽  
Neoadjuvant Chemoradiotherapy ◽  
Wild Type ◽  
Aberrant Expression ◽  
Methods Comparison ◽  
Sox2 Expression ◽  
Before And After

Aim: To determine if neoadjuvant chemoradiotherapy (nCRT) affects p53 and SOX2 expression in esophageal adenocarcinoma (EAC). Materials & methods: Comparison of p53 and SOX2 expression in 100 paired pre- and post-nCRT EAC samples. Results: Aberrant p53 was largely concordant (75/83, 90%), while 13/18 (72%) pre-nCRT samples with wild-type (WT) p53 staining, showed aberrant staining in paired post-nCRT samples. Similarly, 31/45 (69%) with previous WT SOX2 showed SOX2 loss in paired post-nCRT samples, whereas aberrant SOX2 loss was concordant in 50/55 (91%) cases. The prognostic values of both markers regarding survival differ before and after nCRT. Conclusion: Aberrant expression of p53 and SOX2 staining in EAC tissue is unaffected by nCRT. Conversely, the WT-staining pattern frequently changed to aberrant expression.

198 PROGESTERONE, ESTROGEN (ER-α AND ER-β), AND OXYTOCIN RECEPTOR GENE EXPRESSION IN CANINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 248
Author(s):  
A. A. P. Derussi ◽  
A. C. S. Castilho ◽  
R. W. A. Souza ◽  
R. Volpato ◽  
C. R. F. Guaitolini ◽  
...  
Keyword(s):  
Gene Expression ◽  
Relative Abundance ◽  
Hormone Receptors ◽  
Target Genes ◽  
Receptor Gene ◽  
Rna Extraction ◽  
Amplification Efficiency ◽  
Mrna Levels ◽  
Lh Surge ◽  
Total Rna

The aim of this study was to compare the mRNA levels of hormone receptor for progesterone (PR), oestrogen α (ER-α), oestrogen β (ER-β), and oxytocin (OTR) in canine morulae and blastocysts. Ten healthy mature bitches were inseminated based on monitoring vaginal cytology and progesterone concentration. The first insemination was performed on Day 2 after the preovulatory LH surge (progesterone 4 ng mL–1), and the second was performed 48 h later. All females were submitted to ovariohysterectomy (OVH), and the oviduct as well the uterurs were flushed with PBS solution to obtain the embryos. The females were divided into two groups: Group A (n = 5), morulae were collected 8 days after the LH surge and Group B (n = 5), blastocysts were collected 12 days after the LH surge. The pools (n = 10) of embryos (5 embryos/pool) were stored in RNAlater® (Ambion, Life Technologies, USA) at –80°C. The samples were analysed together. The RNA later was removed used PBS calcium free and the total RNA extraction was performed using the Qiagen RNeasy micro-kit (Hildesheim, Germany). Before reverse-transcription (RT) reaction, the total RNA was treated with DNase I Amplification Grade (Invitrogen Life Technologies, Carlsbad, CA, USA). The gene expression of target genes was assessed by real-time RT-qPCR, using SuperScript III for RT and power SYBR Green PCR Master Mix (Applied Biosystems, USA) for cDNA for PCR. The primers for target genes were designed using the software Primer Express® (Applied Biosystems, USA). The gene expression of target genes was normalized by HPRT gene and the relative abundance of mRNA was determined by the ΔΔct method corrected by amplification efficiency using Pffafl’s equation. The means of mRNA relative abundance were compared by t-test. The PR mRNA expression only in blastocysts is similar to the results obtained by Hou et al. (1997) in rat embryos. It is believed that the absence of PR in the early stages of cleavage is due to the indirect action of progesterone by growth factors produced by the maternal reproductive tract (2). Apparently, ER-β action does not occur in the embryo canine phases analysed; however, the action of ER-α seems related to the deployment signal as seen by Hou et al. (1996) in rats. Similarly to findings in the literature, OTR expression decreased in canine embryonic development. This receptor was produced by blastocysts while present in the uterus, which may represent an incidental mechanism to the embryo control of endometrial receptivity, such as also to prevent the development of endometrial luteolytic mechanism. The variation in hormone receptors gene expression in canine embryos can be influencing the transition from morula to blastocyst. In addition, a hormonal influence on these structures can occur in different ways.

213. Regulation of TGFβ superfamily-related genes in the newborn mouse testis by activin

2005 ◽  
Vol 17 (9) ◽  
pp. 82
Author(s):  
S. H. S. Mendis ◽  
C. Brown ◽  
K. L. Loveland
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Kinase Inhibitor ◽  
Rna Extraction ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Pcr Analysis ◽  
Newborn Mouse ◽  
Mrna Levels ◽  
The Impact ◽  
Tgfβ Superfamily

Several members of the transforming growth factor-β (TGFβ) superfamily of ligands are known to influence development of male germ cells. In our ongoing investigations of genetically modified mice with different bioactive levels of activin, we noted that testicular germ cell numbers at birth are increased in the absence of activin.1 In this study we compared expression levels of TGFβ-related genes in mice with 0, 1 or 2 copies of the activin βA subunit gene. Candidate genes were identified through hybridisation of total testis RNA from newborn (Day 0) wild type (WT; 2 copies) and activin βA null mice (KO; 0 copies) to a TGFβ SuperArray Simplicity™ membrane. Densitometric analysis of resulting signals revealed several genes that appeared to differ between genotypes. To validate these findings, pooled testes from newborn WT (2 independent groups), heterozygous (het; 2 groups) and activin βA KO (3 groups) mice were collected for RNA extraction. Quantitative real-time PCR analysis was performed using the Roche Light Cycler, with β-actin mRNA level used as the housekeeping reference. The mRNAs selected for analysis were: inhibin α, p15INK4b, Smad5, insulin-like growth factor 1 (IGF-1), ALK6 (BMPRI-B), and tissue inhibitor of metalloproteinase 1 (Timp1). The inhibin α transcript level was significantly decreased in the activin βA KO animal compared with WT, while the het sample showed an intermediate effect. Both Smad5 and IGF-1 transcripts appeared to increase in the KO animals compared with WT; however, the cyclin-dependent kinase inhibitor, p15INK4b, did not change significantly across the genotypes. Similarly, ALK-6 and Timp1 mRNA levels were also unaffected by genotype. These findings illustrate the impact of graded levels of activin A on specific genes in the fetal testis. (1)Mendis et al. (2004). Reprod. Fertil. Dev. 16(Suppl.), 103.

scholarly journals Warthin tumor of the oropharyngeal minor salivary gland

2018 ◽  
Vol 6 ◽  
pp. 2050313X1881871 ◽  
Author(s):  
Nicole Diaz-Segarra ◽  
Lauren K Young ◽  
Kristin Levin ◽  
William Rafferty ◽  
Joshua Brody ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Parotid Gland ◽  
Minor Salivary Gland ◽  
Previous History ◽  
Salivary Gland Tumor ◽  
Gland Tumor ◽  
Warthin Tumor ◽  
Diagnosis And Management ◽  
History Of ◽  
A Minor

Warthin tumor is the second most common benign salivary gland tumor that classically arises in the parotid gland. It can be synchronous, metachronous, multifocal, bilateral, or unilateral, which complicates diagnosis and management. Rare cases of Warthin tumor of the minor salivary gland are described, but no cases of unilateral, synchronous Warthin tumor involving the parotid and minor salivary gland have been reported. We present a case of Warthin tumor arising from a minor salivary gland in the left oropharynx of a 71-year-old male with a previous history of left parotid Warthin tumor, later determined to be synchronous.

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