Biochemical Genetics最新文献

Breast cancer (BC) remains the most prevalent malignancy among women and a leading cause of cancer-related mortality worldwide. Despite significant advances in recent decades, the molecular mechanisms underlying BC pathogenesis are not yet fully elucidated. Emerging evidence indicates that more than half of the members of the tripartite motif (TRIM) protein superfamily, the majority of which exhibit E3 ubiquitin ligase activity, contribute to BC initiation, progression, and metastasis by exerting functions as either oncoproteins or tumor suppressors. TRIM proteins participate in diverse cellular processes and signaling pathways. In this review, we discuss the specific molecular mechanisms by which TRIM proteins influence BC development, including post-transcriptional modifications, regulation of apoptosis and autophagy, cell cycle control, and metabolic reprogramming of glucose and lipid pathways. A notable feature of TRIM proteins is their engagement in diverse cellular processes and signaling pathways, coupled with their ability to play opposing roles - either promoting or inhibiting BC development - thus reflecting a 'yin and yang' paradigm. Collectively, current data suggest that TRIM genes and their protein products represent promising targets for therapeutic intervention and potential biomarkers for BC prognosis and disease progression.

Membrane-bound O-acyltransferase domain-containing member 7 (MBOAT7) plays an irreplaceable role in lipogenesis and neural development. Over the past decade, a few variants in MBOAT7 have been associated with intellectual disability (ID), and a spectrum of clinical symptoms, including seizures, autism spectrum disorders (ASD), and speech impairment. DNA samples from two of the three affected members of a large Iranian Azeri family with moderate ID and no major dysmorphic features were investigated by whole exome sequencing (WES). Bioinformatic tools were used to identify and evaluate the pathogenicity of the candidate variants. The most likely causative variant was pursued by Sanger sequencing in patients and their relatives. ACMG guidelines and in silico analysis, including molecular modeling, were used to validate the identified variant of uncertain significance (VUS). An in-frame deletion, NM_024298.5: c.37_39 del (p.Leu13del) in the MBOAT7 gene, which had not been previously reported with homozygous genotype, was identified as a possible cause of the clinical conditions in patients. Sanger analysis confirmed the recessive inheritance pattern of this variant in probands and 14 relatives. In-silico modeling of the mutant protein revealed structural changes resulting from removing a highly conserved amino acid. The current study has expanded the MBOAT7 gene variant spectrum and clarified variable phenotypic features to the associated ID criteria. Meanwhile, we have provided insights into the importance of the MBOAT7 protein's first alpha helix in terms of its functionality and solid evidence regarding a VUS's pathogenicity. Additionally, this study presents the oldest recorded case of MBOAT7-related ID in a 51-year-old female to date.

German chamomile (Matricaria chamomilla) is a medicinal herb that promotes improved digestion and reduces insomnia. Although it is widely used worldwide, the mechanism of induction of drug-metabolizing enzymes is unknown. We found that German chamomile extracts induced cytochrome P450 expression at the transcriptional stage. Cyp3a11 expression is decreased at night in wild-type mice, but German chamomile extract induced nocturnal Cyp3a11 and Cyp1a2 expression. German chamomile extract increased the nuclear protein expression of the clock gene BMAL1, which drives and abolishes the rhythm of Cyp3a11 expression. By contrast, German chamomile extract did not significantly alter clock gene expression in the suprachiasmatic nucleus (SCN). Similarly, it did not affect the mRNA expression of the clock genes in the kidneys. Because it did not induce the mRNA expression of ATP-binding cassette (ABC) transporters (Abcb1a, Abcc2, Abcc4, and Abcg2) in the kidney, German chamomile extract had no effect on the transcription of pharmacokinetics-related molecules other than CYPs. German chamomile extract promoted liver-selective nuclear transfer rhythm changes in clock genes and induced the expression of CYPs. This study may help to explain the mechanism of drug interactions associated with chronic German chamomile extract consumption.

Schizophrenia (SCZ) is a deleterious neuropsychological disorder with a worldwide incidence of 1% and unknown etiology. Understanding the role of genetic variants in disease development would enable us to explain the disorder's molecular mechanism and find a more effective prognostic approach. Several studies in various European and East Asian populations have displayed the association of schizophrenia with functional polymorphisms such as rs16944 and rs1143634 that lie within inflammatory pathway genes. This study aimed to evaluate the association of Interleukin-1 beta (IL1B) variants (rs16944, rs1143634) with schizophrenia in the Iranian population for the first time. 565 individuals (240 cases and 325 controls) were recruited. Genotyping was conducted for IL1B single nucleotide polymorphisms (SNPs) (rs16944 and rs1143634) using polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP). In addition, the haplotype analysis was conducted. All statistical analysis was performed using SPSS version 26. Regarding rs1143634 (C > T), T carrier genotypes (CT, TT) compared to CC homozygous genotypes showed a significantly more protective effect (p-value < 0.001). Similarly, concerning the co-dominant model, CT heterozygous genotypes in comparison with homozygous genotypes (CC, TT) illustrated a protective impact regarding schizophrenia (p-value < 0.001). Findings showed a significant difference between cases and healthy controls regarding the rs1143634 (C > T) allele frequency (p-value = 0.025), whereas it determined no considerable difference given rs16944 (p-value = 0.41). Furthermore, in the case of rs16944 (T > C), we found no significant association between case and control groups (p-value = 0.69). Haplotype analysis demonstrated that the C-C (rs1143634-rs16944) haplotype was significantly associated with the risk of schizophrenia (p-value = 0.013). The findings suggest that IL1B rs1143634 (C > T) is significantly associated with SCZ in the Iranian population.

The PI3K-AKT signaling pathway (SP) has been introduced as a key regulatory pathway in cervical cancer (CC). Inhibition of this SP could be a therapeutic strategy in CC. Our previous bioinformatics analysis exhibited that miR-542 could have a dual inhibitory function on this SP by targeting PIK3CB and AKT1 genes with its two arms (-3p and -5p) and proposed it as an appropriate therapeutic target for CC. The current study experimentally investigated the dual inhibitory function of miR-542 on the PI3K-AKT SP. qRT-PCR was performed following transfection of the recombinant pEGFP-C1 vector containing miR-542 precursor into the CaSki and miR-542 overexpression to quantify the expression level of target genes of miR-542 (AKT1 and PIK3CB) as regulators of PI3K-AKT SP and their downstream genes affecting cell proliferation and apoptosis (CDKN1A and BCL2). In addition, the effect of overexpression of miR-542 on cell cycle and apoptosis was examined by flow cytometry using propidium iodide and PE Annexin V/7-AAD staining, respectively. The recombinant cells showed a significant decrease in the expression of AKT1, PIK3CB, and BCL2 genes, and a significant increase in the level of CDKN1A gene expression, simultaneously with the highest overexpression of miR-542 at 48 h post-transfection. Furthermore, the apoptosis was remarkably induced and the cell cycle was arrested in recombinant cells compared to mock cells. miR-542 promoted apoptosis and cell cycle arrest by dual inhibiting the PI3K/AKT SP. It may be introduced as an appropriate target for CC treatment.

Obesity is a multifactorial disorder with a significant genetic component. Variants within the VMAT1 gene have been proposed to influence susceptibility to obesity. This study aimed to assess the association of two single-nucleotide polymorphisms (SNPs), rs2270637 and rs1390938, within VMAT1 with obesity risk in an Iranian population undergoing sleeve gastrectomy. A number of obese patients selected for sleeve gastrectomy were genotyped for rs2270637 and rs1390938. Genotype and allele frequencies were compared using logistic regression under different genetic models (allelic, co-dominant, and recessive). Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to assess the strength of associations. The rs1390938 SNP showed a significant association with obesity in the allelic model, with the A allele conferring protection against obesity (OR = 0.70, 95% CI 0.55-0.88, P = 0.003). In the co-dominant model, individuals with the AA genotype had a significantly reduced risk of obesity compared to those with the GG genotype (OR = 0.20, 95% CI 0.092-0.43, P = 0.000058). Similarly, in the recessive model, the AA genotype remained protective (OR = 0.21, 95% CI 0.099-0.46, P = 0.000087). The rs2270637 SNP also showed significant associations with obesity in co-dominant and recessive models. The GG genotype was protective compared to the CC genotype (OR = 0.11, 95% CI 0.034-0.41, P = 0.001) and compared to the combined GC + CC genotypes (OR = 0.11, 95% CI 0.034-0.40, P = 0.001). Both rs1390938 and rs2270637 polymorphisms in the VMAT1 gene are significantly associated with obesity risk in the studied Iranian cohort. The findings support the role of VMAT1 as a potential genetic susceptibility locus for obesity.

Gemcitabine (GEM) resistance undermines chemotherapy efficacy for pancreatic ductal adenocarcinoma (PDAC), resulting in poor prognosis. Long non-coding RNAs (LncRNAs) participate in various malignant tumors, including PDAC. However, their roles in GEM resistance require further elucidation. Here, we investigated the function of LINC01547 in PDAC progression and chemoresistance. LINC01547 was significantly upregulated in PDAC tissues and cell lines, and its high expression correlated with unfavorable patient outcomes. Silencing LINC01547 dramatically suppressed cellular proliferation, sphere formation capability and enhanced GEM sensitivity of PDAC cells both in vitro and in vivo experiments. Mechanistically, LINC01547 as a competing endogenous RNA that could regulate miR-34a-5p. RNA-sequencing and luciferase reporter analysis demonstrated that miR-34a-5p directly targets MYH9. Additionally, METTL3 mediated m6A modification boosted the RNA stabilization and upregulation of LINC01547. Taken together, these findings indicate that LINC01547 could promote tumor progression and gemcitabine resistance in PDAC via miR-34a-5p/MYH9 axis, highlighting LINC01547 as a potential biomarker and therapeutic target for overcoming chemoresistance in PDAC.

This study investigates the regulatory effects of long non-coding RNA H19 on miR-138-5p and their collective impact on mitochondrial oxidative stress injury in high glucose-exposed cardiomyocytes, while elucidating the underlying molecular mechanisms. The findings aim to establish a theoretical foundation for understanding the pathogenesis of diabetic cardiomyopathy. The expression levels of lncRNA H19, miR-138-5p, and MCU were quantified using RT-qPCR. H9c2 cardiomyocytes were exposed to high glucose (HG, 33 mM) in vitro to establish a diabetic cardiomyopathy (DCM) model. Regulatory targeting relationships between lncRNA H19 and miR-138-5p, as well as between miR-138-5p and mitochondrial calcium uniporter(MCU), were confirmed through dual-luciferase reporter assays. Levels of reactive oxygen species (ROS), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content were quantified to evaluate intracellular oxidative stress in cardiomyocytes. MCU protein expression was analyzed by western blotting. In DCM, H19 and MCU were downregulated; miR-138-5p was upregulated. H19 overexpression increased SOD activity and reduced ROS and MDA levels in HG-treated H9c2 cardiomyocytes. Dual-luciferase assays validated miR-138-5p binding to H19 and MCU 3'UTRs. miR-138-5p overexpression suppressed MCU protein expression. Rescue experiments demonstrated miR-138-5p overexpression or MCU silencing reversed H19-mediated oxidative stress attenuation in HG-stimulated cells. Overexpression of H19 attenuates oxidative stress by modulating the miR-138-5p/MCU axis in DCM, highlighting its potential as a diagnostic biomarker and/or therapeutic target for this condition.

We sought to explore how Tanshinone I (TsI) mediates ferroptosis in femur tissue in a rat model of steroid-induced osteonecrosis of the femoral head (SIONFH). Rats were given lipopolysaccharide and methylprednisolone to develop a rat model of SIONFH and treated with 5 mg/kg or 10 mg/kg TsI or in combination with ferroptosis inhibitor Fer-1. After different treatments, bone parameters (BMD, BV/TV, Tb.N, Tb.Th, and Tb.Sp), the levels of osteoblast markers (RUNX2, BGLAP, and Osteopontin proteins), and ferroptosis markers (SLC7A11, GPX4, and ACSL4) in femur tissues were detected; Additionally, ferroptosis indicators Fe²⁺, MDA, and GSH in femur tissues were detected by corresponding commercial kits. Additionally, this research conducted experiments including TUNEL staining for the cell death rate in femur tissue and immunofluorescence for reactive oxygen species (ROS) detection. The levels of GPX4 (ferroptosis resistance marker), Nrf2, and SLC7A11 through PCR, Western blot, and immunohistochemistry experiments. Furthermore, lentivirus was delivered into SIONFH rats to knock Nrf2 or SLC7A11 down to investigates whether TsI mediated Nrf2/SLC7A11. BMD, BV/TV, Tb.N, and Tb.Th decreased while Tb.SP increased in SIONFH rats, with increased pathological damage to femoral tissue, reductions in expression of osteoblast markers, and increased positive TUNEL signal and cell death rate. Meanwhile, enhanced ferroptosis evidenced by relevant markers was noted in femur tissues. Low- and high-dose TsI treatment attenuated ferroptosis in femoral tissue, improved bone parameters and pathological lesions in SIONFH rats, with the high-dose group demonstrating more pronounced therapeutic effects. Similarly, Fer-1 treatment exerted a comparable protective effect to that of TsI. Mechanistically, low-dose or high-dose TsI treatment up-regulated Nrf2 and SLC7A11 levels, while down-regulation of Nrf2 or SLC7A11 partly compromised the aforementioned impacts of TsI. TsI may alleviate the pathological lesions of SIONFH rats by activating the Nrf2 signaling pathway, thereby promoting SLC7A11 expression and inhibiting ferroptosis in femoral tissue. TsI holds significant potential for therapeutic applications in the treatment of SIONFH.

Enterococci were hospitalized and resistant to numerous antibiotics. The present research aimed to determine the Vancomycin sensitivity by disc and Minimum Inhibitory Concentrations (MICs), also detecting the relationship between Vancomycin resistance genes and the Vancomycin resistance in uropathogenic Enterococcus species. Totally, 150 urine specimens were obtained from patients who attended or admitted to Iraq hospitals. The bacteriological tests besides Vietic and (ddl of E. faecalis, ddl of E. faecium) genes were implemented for the identification of Enterococcus isolates. We indicate 27 Enterococcus isolates, among which 18 (66.7%) were E. faecalis, 4 (14.8%) were E. faecium, while 5 (18.5%) were other isolates belonging to the rest of Enterococcus species. The antibiotic resistance by disk-diffusion technique for Penicillin, Ampicillin, Vancomycin, Norfloxacin, Erythromycin, Ciprofloxacin, Rifampin, Levofloxacin, Chloramphenicol, Fosfomycin, Doxycycline, Minocycline, and Nitrofurantoin was 7 (25,92%), 7 (25,92%), 6 (22.22%), 5 (18.51%), 24 (88.89%), 5 (18.51%), 23 (85.18%), 5 (18.51%), 4 (14.81%), 14 (51.85%), 0 (0%), 3 (11.11%), and 1 (3.70%), respectively. The total isolates were Vancomycin Resistance Enterococci VRE using the MIC technique. Positive results for vanA 23 (85.18%), vanB 11 (40.74%), vanC1 3 (11.11%), vanC2, and vanC3 3 (11.11%). The results revealed that 23 (85.18%) out of VRE isolates possessed the Vancomycin-resistance genes. Although four isolates did not hold van genes, the resistance may be attributed to the occurrence of uncommon genes which not identified by this study; also the efflux pump has a role in resistance. The van genes might be transmitted, so important to monitor antibiotic resistance.