Biochemical Genetics最新文献

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.

Squamous cell carcinoma of the lips and oral cavity (OSCC) contributes to 90% of the nonmelanocytic tumors of these anatomical regions. Liquid biopsy is a new, relatively noninvasive, and effective method for screening, diagnosing, and classifying tumors. Considering the role of integrin β8 subunit (ITGB8) and Myocardial infarction-associated transcript (MIAT) lncRNA in the pathogenesis and progression of different types of cancer, we aimed to evaluate the probable differences in the level of expression of these two biomarkers in control and OSCC cases and evaluate their diagnostic potential as a biomarker in tissue and liquid samples (blood and saliva) biopsy. Tissue, blood, and saliva samples from 60 participants (30 in each study group) were obtained for precise evaluation. Using extracted RNA from the samples and PCR assay, the expression level of these biomarkers was measured in control and case group samples. Parametric and nonparametric tests were equipped for the evaluation of the results. ROC analysis was done to determine the diagnostic potential of these biomarkers. Our results claimed significant differences in the level of expression of ITGB8 and MIAT-lncRNA among our study cohorts' samples. The ROC analysis revealed the valuable potential of these factors in the diagnosis of disease in tissue and blood samples. The results of the analysis for the saliva samples suggest a satisfactory diagnostic value for ITGB8 but nonsignificant diagnostic value for the MIAT-lncRNA. ITGB8 and MIAT-lncRNA expression levels could be considered in the evaluation of suspicious oral cavity lesions, especially in the tissue and blood samples.

Myocardial ischemia-reperfusion injury (MIRI) is a major clinical challenge, marked by metabolic disruptions and cellular damage following the restoration of blood flow after ischemia. During ischemia, the heart shifts from fatty acid metabolism to glucose metabolism, leading to metabolic abnormalities, including reduced intracellular pH, ion disturbances, cell swelling, and apoptosis. Upon reperfusion, fatty acid β-oxidation resumes, becoming the dominant energy source, which induces excessive oxidative stress and aggravates myocardial injury. To explore the role of FTO (Fat mass and obesity-associated protein) in m6A demethylation of MZF1 and its regulation of DECR1, a key enzyme involved in fatty acid β-oxidation, in the context of MIRI. We investigated the molecular mechanisms by which FTO-mediated m6A modification of MZF1 regulates DECR1 expression, leading to enhanced fatty acid oxidation during reperfusion and its contribution to the exacerbation of MIRI. Our findings suggest that FTO-mediated demethylation of MZF1 promotes the expression of DECR1, thereby enhancing fatty acid oxidation. This process intensifies oxidative stress and worsens myocardial injury during ischemia/reperfusion. The FTO-mediated m6A modification of MZF1 represents a critical mechanism in the regulation of fatty acid oxidation and the exacerbation of MIRI. These insights offer potential therapeutic targets to mitigate the harmful effects of reperfusion injury in myocardial infarction.

Background: Lung cancer is a malignant tumor of the bronchial mucosa or gland, the morbidity and mortality increase rapidly, and it is a great threat to human health and life. Propofol is a short-acting intravenous anesthetic, and its effect on lung cancer has been studied, but the mechanism is not thorough.

Methods: The 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU) staining, flow cytometry, and transwell assays were applied to assess the viability, proliferation, apoptosis, and invasion, respectively. The glycolytic analysis was performed using the corresponding kits. The gene expression was evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. The interaction between genes was obtained from the STRING database or ubiquitination analysis. The xenograft tumor mouse models were established to verify the effects of propofol in vivo, and IHC was adopted to detect the gene expression in vivo.

Results: In this study, we found that propofol impeded lung cancer progression and glycolysis. Additionally, propofol curbed the triosephosphate isomerase 1 (TPI1) protein and increased TPI1 ubiquitination modification, meanwhile, propofol exerted inhibitory functions in lung cancer through TPI1. Besides, the protein stability and ubiquitination modification of TPI1 were mediated by ubiquitin-specific peptidase 5 (USP5), and USP5 expedited the progression and glycolysis of lung cancer via TPI1. In the meantime, propofol modulated USP5-regulated functions in lung cancer. In vivo, propofol-inhibited tumor growth by regulating USP5-mediated TPI1.

Conclusion: This study presents propofol/USP5/TPI1 curbing glycolysis metabolism and tumor growth in lung cancer, indicating that propofol-mediated ubiquitination of the target gene may be a new therapeutic target for lung cancer.