Biochemical Genetics最新文献

Non-syndromic hearing loss (NSHL) is a genetically heterogeneous disorder accounting for almost 70% of the total congenital hearing loss. The implementation of rapid advanced sequencing methods has significantly contributed to the correct molecular diagnosis for several rare genetic disorders, including NHSL. Features of two probands with NHSL were clinically and genetically evaluated. One of the affected individuals was subjected to exome sequencing (ES) using standard methods. 3D protein modeling was performed to check the effect of mutation on the protein structure. ES data analysis revealed a homozygous nonsense variant [c.1144A > T; p.Lys382*] within the GPR156 gene (NM_153002.3) associated with rare NSHL. Sanger sequencing supported its recessive segregation within the family. The in silico predictions and 3D protein modeling further affirmed its disease-causing nature. The present study reported a nonsense variant in the GPR156 and its association with NSHL susceptibility, which requires further studies to unveil its key role and disease-related pathophysiology.

The metabolic pathway of aerobic glycolysis in tumor cells has garnered significant attention in tumor research because of its high activation in cancer cells. Previous research conducted by our team has demonstrated that Apolipoprotein M (APOM) exhibits potential as a factor against liver cancer. However, further investigations are needed to elucidate the precise approach and mechanism that are involved in this process. The findings of this study demonstrated that the inhibition of APOM gene expression led to a notable increase in glucose uptake within liver cancer cells, along with increased levels of lactate dehydrogenase A (LDHA) mRNA and protein expression, as well as increased lactate and adenosine triphosphate (ATP) levels (P < 0.05). These alterations in the cellular microenvironment may be associated with a significant increase in the expression level and enzyme activity of the pivotal enzyme hexokinase 2 (HK2) (P < 0.05). Subsequent investigations revealed notable enrichment of the Notch pathway in liver cancer samples exhibiting low expression of the APOM gene. Western blot experiments demonstrated that the inhibition of APOM gene expression triggers the activation of the Notch pathway in liver cancer cells. Furthermore, the administration of a γ-secretase inhibitor (DAPT) successfully mitigated the increase in HK2 levels, glucose uptake, lactate production, and proliferation of liver cancer cells induced by the downregulation of the APOM gene (P < 0.05). In conclusion, diminished APOM expression may facilitate the progression of liver cancer by stimulating the aerobic glycolysis pathway, which is mediated by the Notch signaling pathway.

Lung adenocarcinoma (LUAD) is characterized by its aggressive nature and resistance to treatment. FAM107A is a tumor suppressor gene that has been found to possess inhibitory effects in several cancers, but its role in LUAD remains unclear. This study investigated the role of FAM107A in regulating LUAD cell growth, invasion and aerobic glycolysis and also investigated the potential underlying mechanisms. Our findings revealed that FAM107A is significantly downregulated in LUAD, and its overexpression inhibited LUAD cell growth and invasion. Furthermore, FAM107A overexpression suppressed the anaerobic phase of carbohydrate metabolism in LUAD cells. Mechanistically, FAM107A regulated the CRYAB/PI3K/AKT signaling pathway, thereby inhibiting tumor progression, and similar findings were confirmed in our in vivo mouse model. In conclusion, FAM107A can suppress LUAD progression by regulating the CRYAB/PI3K/AKT pathway and aerobic glycolysis, indicating its potential as therapeutic target for LUAD.

Mounting evidence suggests that circulating microRNAs (miRNAs) hold diagnostic value in various malignancies. To identify circulating miRNAs for the early diagnosis of hepatocellular carcinoma (HCC), we conducted a meta-analysis to evaluate the diagnostic utility of miRNAs in HCC and further validated the results of the meta-analysis. English articles published prior to December 2023 were retrieved from databases including PubMed, Embase, and Web of Science. A random-effects or fixed-effects model was applied depending on the heterogeneity among studies. The pooled sensitivity, specificity, and the area under the summary receiver operating characteristic curve (AUC) were calculated to assess diagnostic accuracy. Additionally, RT-qPCR and receiver operating characteristic (ROC) analyses were employed to further validate the findings. A total of 36 studies were included, involving 3362 patients with HCC and 2150 patients with chronic hepatitis. The pooled sensitivity, specificity, and diagnostic odds ratio were 0.79 (95% CI 0.75-0.82), 0.79 (95% CI 0.73-0.84), and 14 (95% CI 9-22), respectively; the positive and negative likelihood ratios were 4.0 and 0.27, respectively; the area under the curve (AUC) in the summary receiver operating characteristic (ROC) was 0.85 (95% CI 0.82-0.88). Validation indicated a significant upregulation of miR-1246, miR-21, and miR-221 in HCC patients compared to those with chronic hepatitis (P < 0.01), while miR-122 and miR-26a were significantly downregulated (P < 0.01). Moreover, the validation results also demonstrated that serum levels of miR-21, miR-26a, miR-122, miR-221, and miR-1246 exhibit high sensitivity and specificity in the diagnosis of HCC. Circulating miRNAs may be promising biomarkers for HCC diagnosis.

This study aims to establish a genetic risk assessment model based on a score of short tandem repeats (STRs) of polygenic inheritance. A total of 396 children and their biological parents were collected for STR genotyping. The numbers of tandem repeats of two alleles in one STR locus were assumed to be a quantitative genetic strength for disease incidence. The sums of 19 STR loci were considered a quantitative genetic strength per individual. Various thresholds of the STRs between paternal, maternal, and childhood data were recorded. As an exemplar, for thresholds of 25%, the first quarter = 1. All other samples = 0. The consistency rate for heredity (CH) was calculated from the difference in the morbidity of children between parents with and without disease groups. The ratio of observed CH to expected CH was defined as the heredity index (HI). Actual Pedigree data (finger-crossing test) confirmed the accuracy of the STR score. The genetic risk of first-degree relatives could be estimated using easily acquired data (incidence in an unrelated population). Our findings can provide a polygenic genetic model for estimating the incidence and genetic risk of chronic disease in first-degree relatives.

Recent studies highlight the crucial role of microRNAs (miRNAs) in coronary artery disease (CAD). This retrospective study investigated the abundance of miR-432-5p in the serum of CAD patients and explored its role. 252 volunteers were included. The levels of miR-432-5p and Sortilin 1 (SORT1) in the serum of CAD patients and oxidized low-density lipoprotein (ox-LDL)-treated human arterial smooth muscle cells (HASMCs) were quantified via qRT-PCR. The correlation coefficient, clinical diagnostic performance, and risk factors were analyzed with Pearson correlation, receiver operating characteristic (ROC) curve, and binomial logistic regression, respectively. HASMC proliferation, migration, and apoptosis were evaluated using Cell Counting Kit-8 (CCK-8), transwell, and flow cytometry assay, respectively. Potential binding sites between miR-432-5p and SORT1 were predicted with TargetScan and validated through dual-luciferase reporter assay and co-transfection experiments. Serum miR-432-5p was decreased, while SORT1 was elevated in CAD patients and ox-LDL-induced HASMCs. miR-432-5p showed a negative correlation with total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), C-reactive protein (CRP), and Gensini score. miR-432-5p and SORT1 effectively distinguished CAD patients from controls based on ROC analysis. miR-432-5p and SORT1 serve as independent risk predictors. Restoration of miR-432-5p reversed ox-LDL-induced increases in HASMC proliferation and migration and restored apoptosis levels. SORT1 was confirmed as a direct target of miR-432-5p, and its upregulation counteracted the protective effects of miR-432-5p on HASMC under ox-LDL exposure. This study suggests that miR-432-5p protects HASMCs and inhibits coronary artery disease progression by targeting SORT1, positioning both miR-432-5p and SORT1 as potential biomarkers for CAD.

Cisplatin, a platinum-based chemotherapeutic agent, can be used to treat cervical cancer (CC), but cisplatin resistance is increased during the cisplatin treatment. Long non-coding RNA PGM5-AS1 reportedly participates in CC tumorigenesis; however, its role in CC patients with cisplatin resistance has not been revealed. The present aimed to examine the role of PGM5-AS1 in modulating cisplatin resistance in CC. The PGM5-AS1 expression in CC tissues from 29 patients was quantified using quantitative reverse transcription-polymerase chain reaction. The cisplatin-resistant CC cells were constructed by using increasing cisplatin concentrations. The effects of cisplatin resistance interacting with PGM5-AS1 on CC cell malignancy were confirmed by performing Cell Counting Kit 8, colony formation, wound healing, and transwell assays. The key proteins of the Hippo and PI3K-AKT signaling pathways were evaluated by Western blotting. PGM5-AS1 with low expression in CC tissues was correlated to higher International Federation of Gynecology and Obstetrics stage, poor differentiation, lymph node metastasis, and cisplatin resistance. PGM5-AS1 overexpression suppressed the proliferation, migration, and invasion abilities of cisplatin-resistant CC cells. Additionally, PGM5-AS1 overexpression in cisplatin-resistant CC cells could induce the activation of the Hippo signaling pathway and the inactivation of the PI3K-AKT signaling pathway. PGM5-AS1 enhanced the CC cell's sensitivity to cisplatin by activating the Hippo signaling pathway and inactivating the PI3K-AKT signaling pathway. Our study data may provide a novel therapeutic biomarker to overcome cisplatin resistance in CC treatment.

Gastric cancer is associated with high morbidity and mortality rates and seriously threatens human life. Our research aimed to explore the effects of poly (A) binding protein cytoplasmic 1 (PABPC1) on gastric cancer cells and elucidate the underlying mechanisms.

Methods: PABPC1 levels in gastric cancer cell lines were assessed by western blotting and RT-qPCR. Cell viability, apoptosis, invasion, and migration were analyzed using MTT assay, flow cytometry, wound healing assay, and transwell assay, respectively. The levels of apoptosis-related proteins (caspase 3 and cleaved-caspase 3) were determined using the caspase 3 vitality test kit and western blotting. The levels of epithelial-to-mesenchymal transition-related genes (E-cadherin and N-cadherin) in AGS and MGC803 cells were analyzed using western blotting and RT-qPCR. The phosphatidylinositol 3-kinase (PI3K)/AKT pathway was examined using western blot analysis.

Results: PABPC1 expression was enhanced in gastric cancer cells, especially in AGS and MGC803 cells. Our findings indicate that PABPC1 knockdown by siRNA inhibited PABPC1 expression, repressed gastric cancer cell growth, promoted apoptosis, and enhanced cleaved-caspase 3 expression. Functional assays revealed that PABPC1-siRNA blocked the migration and invasion of gastric cancer cells, dramatically promoted E-cadherin expression, and reduced N-cadherin levels. We also found decreased p-PI3K and p-AKT expression, along with decreased p-PI3K/PI3K and p-AKT/AKT in PABPC1-siRNA-treated gastric cancer cells.

Conclusion: PABPC1 silencing in gastric cancer cells inhibited cell proliferation, metastasis, and epithelial-to-mesenchymal transition, partly by repressing the PI3K/AKT signaling pathway activation. This may provide a theoretical basis for gastric cancer therapeutics.

Alzheimer's disease (AD) and mild cognitive impairment (MCI) are a serious global public health problem. The aim of this study was to analyze the key molecular pathological mechanisms that occur in early AD progression as well as MCI. Expression profiling data from brain homogenates of 8 normal volunteers, and 6 patients with prodromal AD who had developed MCI were analyzed, and the data were obtained from GSE12685. Further, overexpression of GABBR2 was achieved in human neuroblastoma cell lines SH-SY5Y and BE(2)-M17 using expression plasmid transfection. GABBR2 was significantly overexpressed in brain tissues of patients with prodromal AD who had developed MCI, as compared to normal brains. Moreover, GABBR2 overexpressing cells showed a significant increase in intracellular Ca²⁺ concentration, a large amount of reactive oxygen species production, a large opening of the mitochondrial permeability transition pore and a significant increase in apoptosis compared with control cells. GABBR2 overexpression was significantly involved in early AD progression and MCI by causing cellular events such as intracellular Ca²⁺ imbalance, oxidative stress, and mitochondrial dysfunction.

Pulmonary hypertension (PH) is a progressive disease characterized by vascular reHypoxiaing, endothelial cell dysfunction, and inflammation. Liver Kinase B1 (LKB1, also known as STK11) is a central regulator of cell polarity and energy homeostasis. However, its specific role and mechanism of action in PH remain unclear. Human pulmonary arterial endothelial cells (hPAECs) were cultured in vitro to establish PH cell Hypoxias under normoxic and hypoxic conditions. The expression of LKB1 was detected by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) and western blotting, and its effect on hPAECs function was investigated by overexpression and inhibition of LKB1. Furthermore, cell proliferation was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis was measured by flow cytometry, inflammatory cytokine secretion was evaluated using enzyme-linked immunosorbent assay (ELISA), and the expression of AMP-activated protein kinase (AMPK) signaling pathway-related proteins was analyzed by western blotting. LKB1 expression was significantly reduced in hypoxia-treated hPAECs compared with that in normoxic controls, and LKB1 overexpression significantly ameliorated the hypoxia-induced decrease in cell proliferation and increase in apoptosis as well as inflammatory factor secretion. The AMPK agonist (GSK621) reversed the dysfunction caused by LKB1 inhibition, indicating that LKB1 regulates hPAECs function through the AMPK signaling pathway. LKB1 plays a protective role in PH by inhibiting hPAECs dysfunction via activation of the AMPK pathway.