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SCD5 mRNA expression in three different cell lines (A) and human tissues. (B) The mRNA expression was measured in HepG2, HEK293T, and SK-N-FI cells, as well as in human liver, kidney, and brain samples. Samples were prepared as described in Section 2. qPCR was performed using GAPDH and SCD5 sequence specific primers as indicated in Section 2. The diagram depicts the results of three independent measurements. Statistical analysis was performed by using the Tukey-Kramer Multiple Comparisons Test. Data are shown as mean values ± S.D. ***: p < 0.001.
Source publication
The combined prevalence of type 1 (T1DM) and type 2 (T2DM) diabetes mellitus is 10.5% worldwide and this is constantly increasing. The pathophysiology of the diseases include disturbances of the lipid metabolism, in which acyl-CoA desaturases play a central role as they synthesize unsaturated fatty acids, thereby providing protection against lipoto...
Contexts in source publication
Context 1
... analysis of the endogenous SCD5 expression in the three cell lines revealed that there were almost completely undetectable traces of SCD5 mRNA in the liver-derived HepG2 cells, while the HEK293T and SK-N-FI cells showed moderate and high mRNA levels of it, respectively ( Figure 2A). The SCD5 promoter constructs and the pCMV-β-gal vector were transiently transfected into HepG2, HEK293T and SK-N-FI cells, and the luciferase activity of the cell lysates was measured and normalized to β-galactosidase 24-30 h after the transfection. ...
Context 2
... analysis of the endogenous SCD5 expression in the three cell lines revealed that there were almost completely undetectable traces of SCD5 mRNA in the liver-derived HepG2 cells, while the HEK293T and SK-N-FI cells showed moderate and high mRNA levels of it, respectively ( Figure 2A). A very similar pattern was observed when performing the comparison of the human tissue RNA samples ( Figure 2B). ...
Context 3
... analysis of the endogenous SCD5 expression in the three cell lines revealed that there were almost completely undetectable traces of SCD5 mRNA in the liver-derived HepG2 cells, while the HEK293T and SK-N-FI cells showed moderate and high mRNA levels of it, respectively ( Figure 2A). A very similar pattern was observed when performing the comparison of the human tissue RNA samples ( Figure 2B). In contrast to the minimal hepatic SCD5 expression, the mRNA levels were one order of magnitude higher in the kidney tissue and two orders of magnitude higher in the brain tissue. ...
Context 4
... very similar pattern was observed when performing the comparison of the human tissue RNA samples ( Figure 2B). In contrast to the minimal hepatic SCD5 expression, the mRNA levels were one order of magnitude higher in the kidney tissue and two orders of magnitude higher in the brain tissue. ...
Citations
... SCD5 is a stearoyl-CoA desaturase involved in monounsaturated fatty acid synthesis. Its biological importance to the developing fetal lung is uncertain, although SNPs in the SCD5 promoter region have been linked to diabetes mellitus [55]. ...
Background
Antenatal steroid (ANS) therapy is given to women at risk of preterm delivery to accelerate fetal lung maturation. However, the benefit of ANS therapy is variable and how maternal and fetal factors contribute to this observed variability is unknown. We aimed to test the degree of concordance in preterm lung function, and correlate this with genomic, transcriptomic, and pharmacokinetic variables in preterm dizygotic twin ovine fetuses.
Methods
Thirty-one date-mated ewes carrying twin fetuses at 123 ± 1 days’ gestation received maternal intramuscular injections of either (i) 1 × 0.25 mg/kg betamethasone phosphate and acetate (CS1, n = 11 twin pairs) or (ii) 2 × 0.25 mg/kg betamethasone phosphate and acetate, 24 h apart (CS2, n = 10 twin pairs) or (iii) 2 × saline, 24 h apart (negative control, n = 10 twin pairs). Fetuses were surgically delivered 24 h after their final treatment and ventilated for 30 min.
Results
ANS-exposed female fetuses had lower arterial partial pressure of carbon dioxide (PaCO2) values than male fetuses (76.5 ± 38.0 vs. 97.2 ± 42.5 mmHg), although the observed difference was not statistically significant (p = 0.1). Only 52% of ANS-treated twins were concordant for lung maturation responses. There was no difference in fetal lung tissue or plasma steroid concentrations within or between twin pairs. Genomic analysis identified 13 single-nucleotide polymorphisms (SNPs) statistically associated with ANS-responsiveness, including in the proto-oncogene MET and the transcription activator STAT1.
Conclusions
Twin fetal responses and ANS tissue levels were comparable with those from singleton fetuses in earlier studies. Twin ovine fetuses thus benefit from ANS in a similar manner to singleton fetuses, and a larger dose of betamethasone is not required. Assuming no difference in input from the placental or maternal compartments, fetal lung responses to ANS therapy in dizygotic twin preterm lambs are dependent on the fetus itself. These data suggest a potential heritable role in determining ANS responsiveness.
... Specifically, we addressed the question whether the alteration of the four nucleotides affected by the polymorphisms could cause a predictable change in the binding probability of any TFs in this region. The in silico TF selection protocol has been published previously 27 . Briefly, for each SNP, the two 41-nucleotide long DNA segments were compared, in which the polymorphic nucleotide was located at position 21. ...
... net/, accessed on 30 June 2022) open-access, nonredundant TF biding profile database was used to predict the potential effect of rs1054411, rs670213, rs2275657 and rs2275656 polymorphisms on TF binding to the SCD1 promoter 59 . The allele-specific effect on TF binding was analyzed as previously described 27 . Briefly, both allelic variants of each SNP were compared pairwise. ...
Overnutrition and genetic predisposition are major risk factors for various metabolic disorders. Stearoyl-CoA desaturase-1 (SCD1) plays a key role in these conditions by synthesizing unsaturated fatty acids (FAs), thereby promoting fat storage and alleviating lipotoxicity. Expression of SCD1 is influenced by various saturated and cis-unsaturated FAs, but the possible role of dietary trans FAs (TFAs) and SCD1 promoter polymorphisms in its regulations has not been addressed. Therefore, we aimed to investigate the impact of the two main TFAs, vaccenate and elaidate, and four common promoter polymorphisms (rs1054411, rs670213, rs2275657, rs2275656) on SCD1 expression in HEK293T and HepG2 cell cultures using luciferase reporter assay, qPCR and immunoblotting. We found that SCD1 protein and mRNA levels as well as SCD1 promoter activity are markedly elevated by elaidate, but not altered by vaccenate. The promoter polymorphisms did not affect the basal transcriptional activity of SCD1. However, the minor allele of rs1054411 increased SCD1 expression in the presence of various FAs. Moreover, this variant was predicted in silico and verified in vitro to reduce the binding of ETS1 transcription factor to SCD1 promoter. Although we could not confirm an association with type 2 diabetes mellitus, the FA-dependent and ETS1-mediated effect of rs1054411 polymorphism deserves further investigation as it may modulate the development of lipid metabolism-related conditions.
... Despite the great attention attributed to SCD1 isoform, few studies on SCD5 isoform have been conducted. The rs3811792 SNP in SCD5 promoter is associated with type 1 and type 2 diabetes [100]. This polymorphism decreases SCD5 promoter activity. ...
Circulating fatty acids (FA) have an endogenous or exogenous origin and are metabolized under the effect of many enzymes. They play crucial roles in many mechanisms: cell signaling, modulation of gene expression, etc., which leads to the hypothesis that their perturbation could be the cause of disease development. FA in erythrocytes and plasma rather than dietary FA could be used as a biomarker for many diseases. Cardiovascular disease was associated with elevated trans FA and decreased DHA and EPA. Increased arachidonic acid and decreased Docosahexaenoic Acids (DHA) were associated with Alzheimer’s disease. Low Arachidonic acid and DHA are associated with neonatal morbidities and mortality. Decreased saturated fatty acids (SFA), increased monounsaturated FA (MUFA) and polyunsaturated FA (PUFA) (C18:2 n-6 and C20:3 n-6) are associated with cancer. Additionally, genetic polymorphisms in genes coding for enzymes implicated in FA metabolism are associated with disease development. FA desaturase (FADS1 and FADS2) polymorphisms are associated with Alzheimer’s disease, Acute Coronary Syndrome, Autism spectrum disorder and obesity. Polymorphisms in FA elongase (ELOVL2) are associated with Alzheimer’s disease, Autism spectrum disorder and obesity. FA-binding protein polymorphism is associated with dyslipidemia, type 2 diabetes, metabolic syndrome, obesity, hypertension, non-alcoholic fatty liver disease, peripheral atherosclerosis combined with type 2 diabetes and polycystic ovary syndrome. Acetyl-coenzyme A carboxylase polymorphisms are associated with diabetes, obesity and diabetic nephropathy. FA profile and genetic variants of proteins implicated in FA metabolism could be considered as disease biomarkers and may help with the prevention and management of diseases.
A certain degree of chromatin openness is necessary for the activity of transcription-regulating regions within the genome, facilitating accessibility to RNA polymerases and subsequent synthesis of regulatory element RNAs (regRNAs) from these regions. The rapidly increasing number of studies underscores the significance of regRNAs across diverse cellular processes and diseases, challenging the paradigm that these transcripts are non-functional transcriptional noise. This review explores the multifaceted roles of regRNAs in human cells, encompassing rather well-studied entities such as promoter RNAs and enhancer RNAs (eRNAs), while also providing insights into overshadowed silencer RNAs and insulator RNAs. Furthermore, we assess notable examples of shorter regRNAs, like miRNAs, snRNAs, and snoRNAs, playing important roles. Expanding our discourse, we deliberate on the potential usage of regRNAs as biomarkers and novel targets for cancer and other human diseases.
Alternative splicing (AS) is a major means of post-transcriptional control of gene expression, and provides a dynamic versatility of protein isoforms. Cancer-related AS disorders have diagnostic, prognostic and therapeutic values. Changes in the expression and AS of human stearoyl-CoA desaturase-5 (SCD5) are promising specific tumor markers, although the transcript variants (TVs) of the gene have not yet been confirmed. Our in silico, in vitro and in vivo study focuses on the distribution of SCD5 TVs (A and B) in human tissues, the functionality of the relevant splice sites, and their modulation by certain single-nucleotide variations (SNVs). An order of magnitude higher SCD5A expression was found compared with SCD5B. This unequal splicing is attributed to a weaker recognition of the SCD5B-specific splicing acceptor site, based on predictions confirmed by an optimized minigene assay. The pronounced dominance of SCD5A was largely modified (rs1430176385_A, rs1011850309_A) or even inverted (rs1011850309_C) by natural SNVs at the TV-specific splice sites. Our results provide long missing data on the proportion of SCD5 TVs in human tissues and reveal mutation-driven changes in SCD5 AS, potentially affecting tumor-associated reprogramming of lipid metabolism, thus having prognostic significance, which may be utilized for novel and personalized therapeutic approaches.