Biochemical Genetics最新文献

Oral cancer, the most prevalent type of cancer in the head and neck region, has an overall five-year survival rate of less than 50%. Key risk factors for its development include tobacco use, alcohol consumption, betel nut chewing, and infections with human papillomavirus (e.g., HPV-16 and HPV-18). While various diagnostic technologies have been developed, some of which have progressed to regulatory-approved in vitro diagnostic systems, there has been no significant improvement in survival rates for patients with oral squamous cell carcinoma despite advancements in surgery, radiotherapy, and chemotherapy. This has prompted the exploration of gene therapy as a novel approach to treating oral cancer. Research indicates that genomic abnormalities and misregulations contribute to both spontaneous and hereditary malignancies. Gene therapy involves the introduction of genetic material into target cells, aiming to minimize harm to surrounding tissues. Various gene therapy techniques, including gene addition therapy, oncolytic virotherapy, suicide gene therapy, excision gene therapy, immunotherapy, and nucleic acid-based therapies, have been investigated both in vitro and in vivo. This review explores these innovative gene therapy strategies, highlighting their potential to address the limitations of conventional treatments and improve outcomes for oral cancer patients.

Musashi-1 (MSI1) has been proposed as a potential prognostic biomarker in prostate cancer (PCa), but its role and underlying molecular mechanisms in PCa progression remain unclear. The mRNA and protein levels of MSI1 and α/β-hydrolase domain 2 (ABHD2) in PCa tissues and cells were examined using qRT-PCR and western blot. Cell proliferation, cycle, apoptosis, and migration were detected by EdU assay, flow cytometry and transwell assay. Glucose uptake and lactate production were assessed to measure cell glycolysis. The interaction between SP1 and PLA2G6 was evaluated using dual-luciferase reporter assay and ChIP assay. MSI1 had increased expression in PCa tissues and cells. MSI1 downregulation could repress PCa cell proliferation, cycle, migration, glycolysis, and enhanced apoptosis. ABHD2 was upregulated in PCa tissues and cells, and MSI1 could bind to ABHD2 promoter region to increase its expression. Knockdown of ABHD2 suppressed PCa cell growth, migration and glycolysis, and ABHD2 overexpression also abolished the effect of MSI1 downregulation on PCa cell progression. Furthermore, interference of MSI1 reduced PCa tumor growth by decreasing ABHD2 expression in vivo. MSI1 facilitated PCa cell proliferation, migration and glycolysis via activating ABHD2 transcription, providing a novel target for PCa treatment.

Breast cancer is the most common invasive cancer diagnosed in females and is also the main cause of cancer-related deaths leading to more than 500,000 deaths annually. The present study aims to identify a promising panel of microRNAs (miRNAs) using bioinformatics analysis, and to clinically validate their utility for diagnosing breast cancer patients with high accuracy in a clinical setting. First, in the in silico phase of our study, using bioinformatics analysis and the data available in the GEO database, miRNAs that were increased in the interstitial fluid of the tumor tissues (differentially expressed miRNAs), were screened and their related target genes were selected. Multimir package of R software was utilized to determine the target genes of the differentially expressed miRNAs (DEMs). The biological functions of discovered genes were analyzed using Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. In order to determine the molecular mechanisms behind important signaling pathways and cellular functions, the protein-protein interaction network was built using STRING and Cytoscape software. After that, in the laboratory phase, the expression level of three candidate miRNAs on the serum samples of 26 breast cancer patients and 26 control, as well as 14 tumor tissue samples and 14 adjacent normal tissue samples, has been investigated by Real-time PCR method. Then sensitivity and specificity of candidate miRNAs were evaluated through the ROC curve analysis. After in silico analysis, we revealed that three miRNAs including miR-4443, miR-572, and miR-150-5p were highly increased in the interstitial fluid of breast cancer patients compared to breast cancer tissues. Moreover, our results revealed that the expression level of miR-4443, miR-572, and miR-150-5p were significantly decreased in the serum of breast cancer patients compare to normal controls. Also, the expression level of miR-4443 and miR-150-5p was significantly decreased in the tumor tissue compared to the adjacent non-tumor tissue. Also, ROC curve analysis showed that these three miRNAs have high sensitivity and specificity for the diagnosis of breast cancer patients. Data analysis was conducted with GraphPad Prism software. Our findings suggest the potential utility of measuring tumor-derived miRNAs in serum as an important approach for the blood-based detection of breast cancer patients. It appears that miR-4443, miR-572, and miR-150-5p may serve as promising diagnostic biomarkers with high sensitivity and specificity. However, it's important to note that further research will be needed to definitively establish the use of these miRNAs as potential biomarkers in clinical practice.

Breast cancer is a heterogeneous tumor with 4 major molecular subtypes. Hormone receptor (HR)-positive and HER2-negative breast cancer accounts for 70% of invasive breast cancers. In our study, we collected 15 original Luminal B breast cancer tissue (LBBC) and paired non-cancerous adjacent tissue (NATs) from patients and performed LC-MS/MS-based label-free quantitative phosphoproteomic analysis. The untargeted metabolomics analysis was also used to determine the differences in metabolic patterns between LBBC and NATs. In addition, an integrative analysis of phosphoproteomics and metabolomics data was performed to investigate regulatory metabolic pathways. The main regulatory proteins were verified by western blot. Phosphoproteomics analysis identified 1385 differentially phosphorylated sites in 785 proteins. The protein kinase A (PKA) and protein kinase C (PKC) families and p70 ribosomal S6 kinase (RPS6K) were strongly activated in LBBC, whereas the cycle-dependent kinases (CDKs) were markedly inhibited. Cancer-specific activation of PI3K-mTORC and Hippo signaling pathways were also highlighted. Metabolomic analysis showed that 223 metabolites were significantly differentially accumulated, including fatty acids (3-hydroxycapric acid; dodecanoic acid; linoleic acid; stearic acid), glycerophospholipids, glycerophosphatidylcholines, and sphingolipids, which were mainly involved in fatty acid oxidation metabolism, sphingolipid metabolism, purine metabolism, and amino acid metabolism pathway. After integrative analysis, we found that the sphingolipid metabolic pathway played the major regulatory role. We also validated 3 phosphorylated proteins (p-YAP, p-SGK1, and p-SGPP2) in the PI3K-mTORC, Hippo signaling pathway, and sphingolipid metabolic pathway, respectively. The present study provides the first integrative phosphoproteome and metabolome profiles of LBBC, mainly involving dysregulation of sphingolipid homeostasis mediated by PI3K-mTORC and Hippo signaling pathways. This study described two phosphorylation pathways and sphingolipid metabolism regulation module for a better understanding of LBBC carcinogenesis and therapy.

Caspase-9 is crucial for initiating apoptosis, and its activity is tightly regulated through various mechanisms, especially phosphorylation by kinases activated by extracellular growth factors, osmotic stress, or during mitosis. Mass spectrometric analyses have shown that residues S302 and S307 in human caspase-9 are prone to phosphorylation. To investigate the effects of phosphorylation at these sites, three phosphomimetic variants of recombinant caspase-9 were created: S302D, S307D, and the combined S302D/S307D variant. The QuickChange method was employed to generate these mutant constructs, which were expressed in Escherichia coli (E. coli) and purified using affinity chromatography. For enzymatic assays, the chromogenic substrate Ac-LEHD-pNA was utilized, and the temperature profiles of enzyme activity were assessed. Computational modeling was used to predict the structures of the mutants, allowing for comparison with the native enzyme. The results indicated that both the S302D and S302D/S307D variants exhibited complete loss of enzyme activity. In contrast, the S307D variant demonstrated a 10-fold increase in the Michaelis constant (K_m) for the substrate and a 4-fold increase in the maximum reaction rate (V_max) compared to the wild-type enzyme. Notably, the k_cat/K_m value for wild-type caspase-9 was three times greater than that of the S307D variant. The optimal temperature for wild-type activity was between 30 and 37 °C, while for the S307D variant, it ranged from 37 to 45 °C. Importantly, the S302 residue is essential for caspase-9 function; introducing a negative charge at this position leads to complete inactivation of the enzyme.

Methotrexate (MTX) pharmacogenetics has been extensively investigated due to the high inter-individual variability in response to treatment. This wide variability can lead to treatment discontinuation or even death. Several genes involved in the pharmacodynamics and pharmacokinetics of MTX have been studied. However, there are still no guidelines for pharmacogenetics-guided MTX dosing. The FPGS rs1544105 and GGH rs3758149 gene polymorphisms were genotyped and their allele frequencies were determined. Their associations with MTX treatment response and toxicity in Uruguayan adults with haematological malignancies receiving high-dose MTX were analyzed. A worldwide systematic review of the association of these gene polymorphisms with response and toxicity to high-dose MTX treatment was also conducted. The allele frequencies of FPGS rs1544105 were 0.54 and 0.46 (C and T, respectively), and of GGH rs3758149 were 0.77 and 0.23 (C and T, respectively). Several associations were found between toxicity (gastrointestinal, hepatic and hematological) and the FPGS rs1544105 T allele (p = 0.01, p < 0.001 and p = 0.04, respectively) and between mucositis and the FPGS TT genotype (p < 0.001). The GGH rs375814 TT genotype was associated with gastrointestinal and hepatic toxicity (p = 0.01 and p < 0.001, respectively). Both the FPGS rs1544105 C allele and the GGH rs3758149 TT genotype were associated with remission (p < 0.001 and p = 0.04, respectively). The systematic review identified 247 publications and finally included 17 research articles. Few consistent data were found due to the lack of homogeneity between study groups.

Anoctamin 6 (ANO6) has been implicated in the oncogenicity of malignancies. However, pan-cancer analysis of ANO6 to fully explore its role in tumors has not been performed and little is reported on its role in melanoma. The ANO6 expression levels, clinical correlation, prognostic significance, mutational profiles, immune infiltration pattern, immune checkpoints, immunomodulatory genes, tumor heterogeneity, and molecular function were explored via systematic bioinformatics analysis and multiple public databases. Subsequently, the biological functions of ANO6 in the pulmonary metastasis of B16F10 melanoma cells in vivo were assessed by experimental validation. Our findings have demonstrated that ANO6 was highly expressed in most cancers and associated with poorer prognosis in cancer patients. A close relationship was observed between ANO6 expression level and clinicopathological characteristics, tumor immunity, and tumor heterogeneity. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that ANO6 was associated with immune response and regulated many cancer-related pathways. Furthermore, a melanoma pulmonary metastasis mice model showed that ANO6 was overexpressed in lung metastasis tissues compared with corresponding normal tissues. Collectively, ANO6 may serve as reliable biomarkers to predict the prognosis for diverse types of cancer and as a prospective marker for melanoma progression.

Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease linked to aging. Mitochondrial dysfunction in circulating T cells, often caused by disruption of mitochondrial DNA (mtDNA), may play a role in age-related conditions like IPF. In our previous study, we found high mtDNA mutational loads in blood lymphocytes from IPF patients, especially in regions critical for mtDNA expression. Since Complex I of the electron transport chain, partly encoded by mtDNA, is essential for energy production, we conducted a preliminary study on its activity. We found significantly reduced Complex I activity (p < 0.001) in lymphocytes from 40 IPF patients compared to 40 controls, which was positively correlated with lung function decline, specifically in functional vital capacity and diffusing capacity for carbon monoxide. These findings indicate that T cell mitochondrial dysfunction is associated with disease progression in IPF. Future work will explore the mechanisms linking T cell mitochondrial disruption with fibrosis, potentially uncovering new therapeutic targets.

Colorectal cancer (CRC) is the most common malignancy of the digestive system. Although research into the causes of CRC's origin and progression has advanced over the past few decades, many details are still not fully understood. Circular RNAs (circRNAs), as a novel regulatory molecule, have been found to be closely involved in various key biological processes in CRC. CircRNAs also have been shown to encode proteins, which could offer new possibilities for therapeutic applications. This ability to produce tumor-specific proteins makes circRNA-based vaccines a potentially valuable approach for targeted cancer treatment. In this review, we summarize recent findings on the various roles of circRNAs in CRC and explore their potential in the development of protein-encoding circRNA vaccines for CRC therapy.

Although secreted frizzled-related protein 4 (SFRP4) has been linked to the development of cardiovascular diseases; it is yet unknown how exactly it functions in arrhythmogenic cardiomyopathy (ACM) remains unclear. Data from the Gene Expression Omnibus (GEO) were used to identify genes that were differentially expressed and linked to ACM. A mouse model known as desmoglein 2 (Dsg2) knockout (Dsg2^-/-) was employed to investigate ACM. Myocardial fibrosis was evaluated by histological analysis, while heart function was evaluated by echocardiography. Angiotensin II (Ang II) was used to stimulate cardiac fibroblasts (CFs) and cause a fibrotic phenotype. The ability of CFs to migrate was evaluate using a wound healing assay. Gene Set Enrichment Analysis (GSEA) was used to do an enrichment study of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The levels of SFRP4, transforming growth factor beta receptor 2 (TGFBR2), TGF-β2, and Smad family member 3 (Smad3) were assessed using quantitative real-time PCR and Western blot. Our findings show that SFRP4 is highly expressed in Dsg2^-/- mice. SFRP4 knockdown markedly reduced myocardial fibrosis, ventricular compliance, and cardiac dilation in Dsg2^-/- mice. The level of SFRP4 was higher in CFs treated with Ang II, andSFRP4 inhibition markedly decreased the migration of Ang II-induced CFs. Moreover, SFRP4 activates the TGF-β signaling pathway, with SFRP4 knockdown resulting in a significant decrease in the expression levels of TGF-β2, TGFBR2, and Smad3 in Dsg2^-/- mice. In summary, SFRP4 knockdown reduced cardiac fibrosis in ACM by inhibiting the TGF-β signaling pathway.