Ekspresja genów receptorów melatoninowych i genów związanych z regulacją ich aktywności w gruczolakoraku endometrium

Objectives: The aim of the study was to evaluate transcription activity of melatonin receptors and genes associated with regulation of their activity in endometrial adenocarcinoma to identify probable diagnostic and prognostic molecular markers. Material and methods: The material included endometrial adenocarcinoma tissue samples of histopathological grades G1, G2, G3, and normal endometrium. The molecular analysis was performed on 37 patient samples. Total RNA was extracted and used for the microarray HG-U133A analysis. Among 22 283 ID mRNA, only entities of genes associated with regulation of melatonin receptors activity were selected. qRT-PCR was employed for validation, what allowed to compare melatonin receptor genes activation in endometrial cancer tissues to the normal endometrium. Results: The results of the microarray experiments showed that only 18 ID mRNA were differential in endometrial cancer samples as compared to the control at p-value1.5. These genes were identified as differentially expressed in grade G2- ASMTL, GNA11, PER2, PTGDS and in grade G3- GNAI2, GNA11. Silencing of RGS4 encoding RGP4, which regulates signal transmission by G protein, was observed in all cancer groups, independently of the histopathological grade. Conclusions: The profile expression of genes associated with regulation of melatonin receptors activity was different and dependent on the histopathological grade of endometrial cancer and can be an additional diagnostic and prognostic marker. Statistically significant was the down-regulation of melatonin biosynthesis genes (ASMTL) and melatonin signal transmitters (GNA11, GNA12, RTGSCel pracy: Celem pracy była ocena aktywności transkrypcyjnej genów kodujących receptory melatoninowe oraz genów zaangażowanych w ich regulację w gruczolakoraku endometrium pod kątem poszukiwania molekularnych markerów diagnostycznych i prognostycznych tego nowotworu. Materiał i metody: Materiał do badań stanowiły wycinki gruczolakoraka endometrium typu endometrioidalnego w stopniu histologicznego zróżnicowania G1, G2, G3 oraz endometrium prawidłowego. Badania molekularne wykonano u 37 kobiet. Po izolacji RNA z tkanek techniką mikromacierzy oligonukleotydowych HG-U133A (Affymetrix) spośród 22 283 ID mRNA wyznaczono profil stężenia mRNA genów związanych z aktywnością receptorów melatoninowych. Metodą qRT-PCR oceniono zmiany aktywności transkrypcyjnej genów receptorów melatoninowych w wycinkach gruczolakoraka w porównaniu do endometrium prawidłowego. Wyniki: Analiza transkryptomów metodą mikromacierzy ekspresyjnych (Affymetrix) pozwoliła na wytypowanie 18 ID mRNA dla genów związanych z aktywnością receptorów melatoninowych różnicujących raka endometrium od kontroli, przy założeniu, że wartość p1,5. Wśród genów specyficznie różnicujących raka są: w stopniu G2- ASMTL, GNA11, PER2, PTGDS oraz w stopnia G3- GNA12, GNA11. Wyciszenie genu kodującego białko RGS4, regulujące czas i nasilenie przekazywania sygnału przy udziale białka G (negatywnego regulatora) obserwowano we wszystkich wycinkach raka, niezależnie od stopnia zróżnicowania histologicznego. Wnioski: Profil stężenia mRNA genów uczestniczących w regulacji aktywności biologicznej receptorów melatoninowych zależy od stopnia zróżnicowania histologicznego gruczolakoraka endometrium i może stanowić uzupełniający marker diagnostyczny i prognostyczny raka endometrium. Istotne statystycznie obniżenie ekspresji genów zaangażowanych w proces biosyntezy melatoniny (ASMTL) oraz genów kodujących białka związane z transdukcją sygnału receptorów melatoninowych (GNA11, GNA12, RTGS4, HTR2B i GNRH2), mogą stanowić obiecujące ogniwo w podjęciu nowych terapeutycznych rozwiązań w leczeniu gruczolakoraka endometrium

Vitamin D Receptor gene (VDR) transcripts in bone, cartilage, muscles and blood and microarray analysis of vitamin D responsive genes expression in paravertebral muscles of Juvenile and Adolescent Idiopathic Scoliosis patients

Abstract Background VDR may be considered as a candidate gene potentially related to Idiopathic Scoliosis susceptibility and natural history. Transcriptional profile of VDR mRNA isoforms might be changed in the structural tissues of the scoliotic spine and potentially influence the expression of VDR responsive genes. The purpose of the study was to determine differences in mRNA abundance of VDR isoforms in bone, cartilage and paravertebral muscles between tissues from curve concavity and convexity, between JIS and AIS and to identify VDR responsive genes differentiating Juvenile and Adolescent Idiopathic Scoliosis in paravertebral muscles. Methods In a group of 29 patients with JIS and AIS, specimens of bone, cartilage, paravertebral muscles were harvested at the both sides of the curve apex together with peripheral blood samples. Extracted total RNA served as a matrix for VDRs and VDRl mRNA quantification by QRT PCR. Subsequent microarray analysis of paravertebral muscular tissue samples was performed with HG U133A chips (Affymetrix). Quantitative data were compared by a nonparametric Mann Whitney U test. Microarray results were analyzed with GeneSpring 11GX application. Matrix plot of normalized log-intensities visualized the degree of differentiation between muscular tissue transcriptomes of JIS and AIS group. Fold Change Analysis with cutoff of Fold Change ≥2 identified differentially expressed VDR responsive genes in paravertebral muscles of JIS and AIS. Results No significant differences in transcript abundance of VDR isoforms between tissues of the curve concavity and convexity were found. Statistically significant difference between JIS and AIS group in mRNA abundance of VDRl isoform was found in paravertebral muscles of curve concavity. Higher degree of muscular transcriptome differentiation between curve concavity and convexity was visualized in JIS group. In paravertebral muscles Tob2 and MED13 were selected as genes differentially expressed in JIS and AIS group. Conclusions In Idiopathic Scolioses transcriptional activity and alternative splicing of VDR mRNA in osseous, cartilaginous, and paravertebral muscular tissues are tissue specific and equal on both sides of the curve. The number of mRNA copies of VDRl izoform in concave paravertebral muscles might be one of the factors differentiating JIS and AIS. In paravertebral muscles Tob2 and Med13 genes differentiate Adolescent and Juvenile type of Idiopathic Scoliosis.</p

Evaluation of Polymeric Matrix Loaded with Melatonin for Wound Dressing

The development of scaffolds mimicking the extracellular matrix containing bioactive substances has great potential in tissue engineering and wound healing applications. This study investigates melatonin—a methoxyindole present in almost all biological systems. Melatonin is a bioregulator in terms of its potential clinical importance for future therapies of cutaneous diseases. Mammalian skin is not only a prominent melatonin target, but also produces and rapidly metabolizes the multifunctional methoxyindole to biologically active metabolites. In our methodology, chitosan/collagen (CTS/Coll)-contained biomaterials are blended with melatonin at different doses to fabricate biomimetic hybrid scaffolds. We use rat tail tendon- and Salmo salar fish skin-derived collagens to assess biophysical and cellular properties by (i) Fourier transform infrared spectroscopy—attenuated total reflectance (FTIR–ATR), (ii) thermogravimetric analysis (TG), (iii) scanning electron microscope (SEM), and (iv) proliferation ratio of cutaneous cells in vitro. Our results indicate that melatonin itself does not negatively affect biophysical properties of melatonin-immobilized hybrid scaffolds, but it induces a pronounced elevation of cell viability within human epidermal keratinocytes (NHEK), dermal fibroblasts (NHDF), and reference melanoma cells. These results demonstrate that this indoleamine accelerates re-epithelialization. This delivery is a promising technique for additional explorations in future dermatotherapy and protective skin medicine