Coronary heart disease (CHD) is driven by endothelial dysfunction and chronic vascular inflammation. hsa-miR-2110 (miR-2110) has been associated with adverse cardiovascular outcomes, but its mechanistic role in CHD remains unclear. In this study, miR-2110 expression was quantified in peripheral blood from CHD patients and healthy controls. Functional effects were assessed in EA.hy926 endothelial cells following lentiviral overexpression of miR-2110. The target gene TRAF3 was identified by RNA sequencing, bioinformatic analysis, and validated by dual-luciferase reporter assays, RT-qPCR, and Western blotting. TRAF3 expression was further evaluated in CHD patient blood samples and in atherosclerotic lesions from ApoE−/− mice fed a high-fat diet. miR-2110 was significantly downregulated in CHD patients. Overexpression of miR-2110 in endothelial cells impaired proliferation and migration, induced S-phase arrest, reduced apoptosis, and promoted cellular senescence. TRAF3 was confirmed as a direct target of miR-2110. TRAF3 was significantly upregulated in CHD patients. In ApoE−/− mice, TRAF3 protein expression was increased in atherosclerotic lesions, predominantly within the tunica intima. Pathway enrichment predicted NF-κB–related signaling among the enriched pathways potentially associated with the miR-2110/TRAF3 axis. Together, our findings suggest that the miR-2110/TRAF3 axis represents a novel regulatory pathway involved in CHD, potentially relevant to endothelial dysfunction and inflammatory signaling.

Background Vascular aging involves structural remodeling, impaired vascular homeostasis, and cellular senescence. Iron dyshomeostasis and ferroptosis may contribute to these changes, but the role of sirtuin 3 (SIRT3)–glutathione peroxidase 4 (GPX4) signaling in endothelial senescence remains unclear. Objective To investigate the association of the SIRT3–GPX4 axis with ferroptosis-related endothelial senescence and vascular aging. Methods Middle-aged mice were subjected to iron overload, and endothelial senescence was induced in EA.hy926 cells using D-galactose. Ferrostatin-1 treatment and SIRT3 overexpression were used to assess senescence, ferroptosis-related alterations, and mitochondrial dysfunction. Co-immunoprecipitation and acetyl-lysine immunoblotting were performed to examine SIRT3–GPX4 association and GPX4 acetylation. Results Iron overload aggravated elastic fiber disruption, collagen deposition, and senescence-associated changes in mouse aortas, accompanied by increased acyl-CoA synthetase long-chain family member 4 (ACSL4) and reduced GPX4 and SIRT3 expression. Ferrostatin-1 partially attenuated these alterations. In D-galactose-treated endothelial cells, reduced GPX4 expression, lipid reactive oxygen species accumulation, increased intracellular Fe²⁺, and mitochondrial dysfunction were observed. Ferrostatin-1 alleviated lipid peroxidation and mitochondrial abnormalities without restoring SIRT3 expression. SIRT3 overexpression attenuated ferroptosis- and senescence-associated changes, improved mitochondrial function, increased GPX4 abundance, and was associated with reduced GPX4 acetylation. Reciprocal co-immunoprecipitation supported an endogenous association between SIRT3 and GPX4. Conclusions Reduced SIRT3 expression was associated with GPX4 acetylation, ferroptosis-related stress, endothelial senescence, and vascular aging. These findings support a potential role for SIRT3–GPX4-associated regulation in linking iron dyshomeostasis to vascular aging.

Glucagonoma, a rare neuroendocrine tumor, lacks targeted treatment drugs. Excessive secretion of glucagon is the main cause of its clinical syndrome. To explore targeted therapeutic drugs that can inhibit glucagon secretion and tumor proliferation, we investigated the effect of Trametenolic Acid (TA) on mouse pancreatic alpha TC1 clone 6 (αTC1-6) cells and its regulatory role in the PI3K/AKT signaling pathway. Cell viability of αTC1-6 cells was assessed via the MTT assay. Glucagon content in cell culture supernatants was measured using an Enzyme-Linked Immunosorbent Assay (ELISA). Autophagic vacuoles were visualized through Monodansylcadaverine (MDC) staining. The expression of autophagy-related proteins including Atg7, LC3 Ⅱ and PI3K/AKT signaling pathway-related proteins mTOR and FoxO1 were determined by Western blot. The results showed that the proliferation of αTC1-6 cells was significantly inhibited by TA in a dose- and time-dependent manner, and the IC50 was 140.71, 26.77 and 1.99 μM after treatment of 12, 24, and 48 h, respectively. The secretion of glucagon was significantly inhibited by TA. The MDC staining results showed that the fluorescent labeled autophagic vesicles in the TA group were increased. The Western blot results showed that the expression of Atg7 and LC3 Ⅱ was promoted by TA in a dose-dependent manner, the phosphorylation of PI3K, AKT, mTOR and FoxO1 was significantly inhibited, and the expression of FoxO1 protein was increased. These results demonstrated that TA can inhibit glucagon secretion, induce autophagy, and suppress cell proliferation in αTC1-6 cells. The mechanism may be associated with the PI3K/AKT signaling pathway.

miR-27a-3p is a multifunctional miRNA that plays a critical role in the process of angiogenesis. However, its specific effect on coronary heart disease (CHD), particularly on the regulation of downstream molecules and the resulting impact on endothelial cell injury, has not yet been fully elucidated. This study aimed to explore the relationship between miR-27a-3p and CHD and its underlying mechanical molecular pathways in CHD patients and modeled endothelial cells with techniques such as RT-qPCR, RNA sequencing and bioinformatics. Consequently, the expression of miR-27a-3p was significantly decreased in CHD patients. In endothelial cells, overexpression of miR-27a-3p was observed to decrease malonaldehyde, gamma H2A histone family member X and interleukin 6 while increased superoxide dismutase, thus reduced endothelial injury and senescence. RNA sequencing and bioinformatics revealed glutamate ionotropic receptor NMDA type subunit 2D (GRIN2D) as a target gene of miR-27a-3p, and dual luciferase assays confirmed the direct binding of miR-27a-3p to the 3’UTR of GRIN2D. Subsequent validation experiments demonstrated that miR-27a-3p inhibited the protein expression of GRIN2D and PKC and suppressed the activation of the MAPK/ERK signaling pathway by reduced downstream MEK and ERK phosphorylation, leading to enhanced endothelial apoptosis. In conclusion, miR-27a-3p played a crucial role in regulating endothelial cell dysfunction which may trigger coronary atherosclerosis and CHD by targeting GRIN2D in the PKC/MEK/ERK signaling pathway.

To investigate how aging hallmarks exert roles in the age-related disease of coronary artery disease (CAD). R software and the GEO2R online tool identified differentially expressed genes (DEGs) and differentially expressed microRNAs (DEMis) in CAD microarray datasets from the Gene Expression Omnibus. Genes common to target genes of DEMis, DEGs, and an aging gene list from Human Aging Genomic Resources were then identified and analyzed for protein-protein interactions and functional and pathway enrichment. An miR-mRNA network was constructed using Cytoscape. Receiver operating characteristic curve analysis assessed the diagnostic utility of DEMis in CAD. The expression of two DEMis from a CAD cohort was employed to validate the findings. An aging hallmark gene set, comprising 18 genes, was delineated, with the hub gene TP53 established through protein-protein interaction and microRNA-mRNA networks. Within the microRNA-mRNA network, two DEMis (hsa-miR-423-5p and hsa-miR-564) potentially regulated TP53, rendering them potential CAD biomarkers, as indicated by their area under the curves (AUC) surpassing 0.6. Validation experiments corroborated an AUC of 0.7002 for hsa-miR-423-5p and 0.7261 for hsa-miR-564, highlighting its protective association with CAD. Combining hsa-miR-423-5p, hsa-miR-564, total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), white blood cells (WBC) achieved an area under the receiver operating characteristics curve of 0.783. A CAD-associated gene set was identified, with TP53 as the central hub. Hsa-miR-564 emerged as a potential protective factor against CAD.

Renal fibrosis (RF) is a typical pathological presentation of end-stage chronic kidney disease (CKD) and autosomal dominant polycystic kidney disease (ADPKD). However, the precise regulatory mechanisms governing this re-expression process remain unclear. Differentially expressed microRNAs (miRNAs) associated with RF were screened by microarray analysis using the Gene Expression Omnibus (GEO) database. The miRNAs upstream of the genes in question were predicted using the miRWalk database. The miRNAs involved in the two GEO data sets were intersected to identify key miRNAs; their regulatory pathways were investigated using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Subsequently, the effects and the underlying mechanisms of target miRNA on RF were examined in a unilateral ureteral obstruction (UUO)-induced mice renal fibrotic model and a transforming growth factor-beta 1 (TGF-beta 1)-induced tubular epithelium (HK-2) fibrotic cell model. In total, 109 and 32 differentially expressed miRNAs were identified in the GSE133530 and GSE80247 data sets, respectively. GREM1 was identified as a hub gene, where its 2196 upstream miRNAs were predicted; miR-574-5p was found to be downregulated and closely related to fibrosis after data set intersection and enrichment analyses, thus was selected for further investigation. A differential expression heatmap (GSE162794) showed that miR-542-5p was downregulated. The expression of GREM1 mRNA was upregulated, whereas that of miR-542-5p was downregulated in UUO mice and fibrotic HK-2 cells as compared with the relevant controls. The binding site of miR-542-5p was predicted at the 3'UTR region of GREM1 and was confirmed by subsequent dual luciferase reporter gene assay. Western blot analysis showed that Gremlin-1 and Fibronectin were significantly upregulated after induction of TGF-beta 1; when miR-542-5p was overexpressed or GREM1 mRNA was interfered, the upregulations of Gremlin-1 and Fibronectin were significantly reduced. Our research demonstrates that miR-542-5p plays a critical role in the progression of RF, and thus may be a promising therapeutic target for CKD and ADPKD.

In search of an effective therapeutic target for bladder urothelial carcinoma (BLCA), the present study aimed to investigate the expression of cyclin B1 (CCNB1) and its putative mechanism in BLCA. BLCA sequencing data from Gene Expression Omnibus and The Cancer Genome Atlas were used to analyze expression of CCNB1 mRNA and high CCNB1 expression had a poorer prognosis compared with those with low expression. Immunohistochemistry (IHC) samples collected from the Human Protein Atlas database were analyzed for CCNB1 protein expression. Short hairpin (sh) CCNB1-transfected BLCA T24 and 5637 cells were used to investigate the effects of CCNB1 and inhibit the proliferation, migration and invasion of BLCA cells, affect the cell cycle distribution and promote apoptosis of 5637 cells. A sh-CCNB1 BLCA chicken embryo chorioallantoic membrane (CAM) transplantation model was established to observe the impacts of sh-CCNB1 on the tumorigenesis of BLCA in vivo. Analysis of sequencing data showed that CCNB1 mRNA was significantly elevated in tumor and BLCA compared with normal tissues [standardized mean difference (SMD)=1.21; 95% CI: 0.26-2.15; I²=95.9%]. IHC indicated that CCNB1 protein was localized in the nucleus and cytoplasm and was significantly increased in BLCA tumor tissues. The in vitro tests demonstrated that proliferation of T24 and 5637 cells transfected with sh-CCNB1 was significantly inhibited and cell migration and invasion ability were significantly decreased. sh-CCNB1 decreased the percentage of T24 cells in G0/G1, 5637 cells in the G0/G1 phase and S phase and increased percentage of 5637 cells in the G2/M phase and increased early apoptosis of 5637 cells. The in vivo experiments demonstrated that the mass of transplanted tumors was significantly decreased compared with the control group following silencing of CCNB1. The present results suggested that CCNB1 was involve in the development and prognosis of BLCA and silencing of CCNB1 may be a promising targeted therapy for BLCA.

目的:探讨细胞周期蛋白B1(CCNB1)在肝癌(HCC)中的表达及沉默CCNB1基因对肝癌细胞的影响.方法:利用Gene Expression Omnibus(GEO)和 Genotype-Tissue Expression(GTEx)数据库中的HCC测序数据,对CCNB1 mRNA的表达水平及其与预后的相关性进行综合分析.在Huh7、SK两种HCC细胞系中沉默CCNB1(sh-CCNB1)检测其对HCC细胞生长、迁移、侵袭和细胞周期的影响.结果:测序数据分析发现,与正常组织比较,CCNB1 mRNA表达水平在多种肿瘤组织中显著升高,在HCC组织中也显著升高(SMD=1.69,95％CI:1.36～2.02,I2=96.2％).在Huh7、SK细胞系中下调CCNB1表达结果发现,相对于对照组(sh-NC组),sh-CCNB1组细胞增殖受到明显抑制,细胞迁移和侵袭能力均显著下降;与sh-NC组相比,sh-CCNB1使Huh7细胞G0/G1期占比增加(P＜0.05),S期减少(P＜0.05);使SK细胞S期占比减少(P＜0.000 1)、G2/M期占比显著增多(P＜0.01).结论:CCNB1在HCC中高表达,且沉默CCNB1抑制肝癌细胞的生长.

文章立足于新医科背景,在深入剖析广西医科大学临床医学教学改革班呼吸系统基础与疾病教学模块整合前后所遇主要问题及原因的基础上,系统阐述了呼吸系统基础与疾病教学团队在课程整合及优化中所采取的系列教学改革措施及课程建设取得的初步成效.

Coronary heart disease (CHD) is a prevalent global cause of death. Research suggests that circular RNAs (circRNAs) play a role in the development of CHD. In this study, we investigated the expression of hsa_circRNA_0000284 in peripheral blood leukocytes (PBLs) obtained from a cohort of 94 CHD patients aged over 50 years, as well as 126 age-matched healthy controls (HC). An in vitro inflammatory and oxidative injury cell model that simulates CHD was used to evaluate changes in hsa_ circRNA _0000284 under stress. CRISPR/Cas9 technology was used to evaluate changes in hsa_circRNA_0000284 expression. An hsa_ circRNA_0000284 overexpression and silencing cell model was used to analyze the biological functions of hsa_circRNA_0000284. Bioinformatics, qRT-PCR, viral transfection technology, and luciferase assays were used to evaluate the potential hsa_circRNA_0000284/miRNA-338-3p/ETS1 axis. Western blotting analysis was performed to detect protein expression. Herein, PBLs from CHD patients exhibited downregulation of hsa_circRNA_0000284 expression. Exposure to oxidative stress and inflammation can induce damage to human umbilical endothelial cells, resulting in the downregulation of hsa_circRNA_0000284 expression. The expression of hsa_circRNA_0000284 in EA-hy926 cells was significantly reduced after the AluSq2 element of hsa_circRNA_0000284 had been knocked out. The expression of hsa_circRNA_0000284 affected proliferation, cycle distribution, aging, and apoptosis in EA-hy926 cells. Consistent with the results of cell transfection experiments and luciferase assays, Western blotting showed that hsa_circRNA_0000284 plays a role in the regulation of hsa-miRNA-338-3p expression. Subsequently, hsa-miRNA-338-3p was found to be involved in the regulation of ETS1 expression.Communicated by Ramaswamy H. Sarma.

Abstract The progression of prostate cancer (PCa) leads to poor prognosis. However, the molecular mechanism of PCa is still not completely clear. This study aimed to elucidate the important role of centromere protein A (CENPA) in PCa. Large numbers of bulk RNA sequencing (RNA‐seq) data and in‐house immunohistochemistry data were used in analysing the expression level of CENPA in PCa and metastatic PCa (MPCa). Single‐cell RNA‐seq data was used to explore the expression status of CENPA in different prostate subpopulations. Enrichment analysis was employed to detect the function of CENPA in PCa. Clinicopathological parameters analysis was utilised in analysing the clinical value of CENPA. The results showed that CENPA was upregulated in PCa (standardised mean difference [SMD] = 0.83, p = 0.001) and MPCa (SMD = 0.61, p = 0.029). CENPA was overexpressed in prostate cancer stem cells (CSCs) with androgen receptor (AR) negative compared to epithelial cells with AR positive. CENPA may influence the development of PCa through affecting cell cycle. Patients with nodal metastasis had higher expression level of CENPA. And patients with high CENPA expression had poor disease‐free survival. Taken together, Overexpression of CENPA may influence the development of PCa by regulating cell cycle and promoting metastasis.

Aim To explore the correlation between protein tyrosine kinase 7(PTK7)and coronary heart disease(CHD)and its diagnostic value.Methods Target genes were obtained through the Gene Expression Omnibus(GEO)database.StataSE15 was used to find the total standardized mean difference(SMD)of PTK7 and plot the sum-mary receiver operating characteristic(SROC)curve.Next,reverse transcription quantitative polymerase chain reaction was used to verify the expression of PTK7 in CHD and non-CHD population samples and search for CHD-related single-cell RNA sequencing data from GEO to analyze the expression of PTK7 in different cells.The upstream transcription factor(TF)of PTK7 was predicted by the Cistrome Data Browser database.Moreover,enrichment analysis of the gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed on the differentially co-expressed genes.Results By calculating the SMD of PTK7,it was found that PTK7 was highly expressed in the peripheral blood leukocytes(PBL)of patients with CHD(total SMD=0.81,95％confidence interval=0.17～1.45).The population sample vali-dation confirmed the above results.When SROC was plotted,the area under the curve(AUC)was 0.79,indicating that PTK7 has the ability to distinguish between CHD and non-CHD.The single-cell RNA sequencing results showed that the expression ratio of PTK7 was relatively low in different cells of normal peripheral blood.In addition,potential upstream TFs of PTK7 were predicted through the ChIP-seq database,where it was found that IRAKI,SNAI2 may be positive up-stream TFs of PTK7,and EP300,NIPBL may be negative upstream TFs of PTK7.Conclusion Highly expressed PTK7 in PBL is positively correlated with the pathogenesis of CHD,demonstrating that PTK7 has definite diagnostic value on CHD.

ObjectsTo evaluate the hsa_circ_0001445 level in peripheral blood leukocytes of patients with coronary heart disease (CHD) and its related clinical factors, and predict its circRNA-miRNA-mRNA regulatory network in CHD pathogenesis via bioinformatics analysis. MethodsPeripheral blood leukocytes were isolated from the whole blood samples of 94 CHD patients (aged 65.96 +/- 9.78 years old) and 126 healthy controls (aged 60.75 +/- 8.81 years old). qRT-PCR was used to quantify the expression level of circRNA and subsequently analyze its association with CHD clinical parameters. Via bioinformatics algorithm and GEO datasets, differential miRNA expression was evaluated using the Limma package. A miRNA-mRNA regulatory network was predicted by cyTargetLinker. ClusterProfiler was employed to perform functional enrichment analysis of the circRNA network to investigate its role in CHD pathogenesis. ResultsThe expression of hsa_circ_0001445 in peripheral blood leukocytes of CHD patients was downregulated compared with that of healthy controls. Positive correlations were evident between hsa_circ_0001445 expression level and the levels of hemoglobin, triglycerides, high- and low-density lipoprotein cholesterol. A significant negative correlation was also found between hsa_circ_0001445 expression level and age and the neutrophil level. Low expression of hsa_circ_0001445 exhibited a discriminatory ability between CHD patients and healthy controls with a sensitivity of 67.5% and a specificity of 76.6% (p < 0.05). By bioinformatics analysis, 405 gene ontology terms were identified. The Kyoto Encyclopedia of Genes and Genomes terms focused principally on the PI3K-Akt signaling pathway. hsa_circ_0001445 was associated with the expression of three miRNAs that may regulate 18 genes involved in KEGG processes: hsa-miR-507, hsa-miR-375-3p, and hsa-miR-942-5p. ConclusionThe hsa_circ_0001445 level in peripheral blood leukocytes may serve as a biomarker for CHD diagnosis. Our work on circRNA-miRNA-mRNA networks suggests a potential role for hsa_circ_0001445 in CHD development.

Background: To date, the clinical management of advanced hepatocellular carcinoma (HCC) patients remains tough and the mechanisms of E2F transcription factor 1 (E2F1) underlying HCC are obscure. Materials and Methods: Our study integrated datasets mined from several public databases to comprehensively understand the deregulated expression status of E2F1. Tissue microarrays and immunohistochemistry staining was used to validate E2F1 expression level. The prognostic value of E2F1 was assessed. In-depth subgroup analyses were implemented to compare the differentially expressed levels of E2F1 in HCC patients with various tumor stages. Functional enrichments were used to address the predominant targets of E2F1 and shedding light on their potential roles in HCC. Results: We confirmed the elevated expression of E2F1 in HCC. Subgroup analyses indicated that elevated E2F1 level was independent of various stages in HCC. E2F1 possessed moderate discriminatory capability in differentiating HCC patients from non-HCC controls. Elevated E2F1 correlated with Asian race, tumor classification, neoplasm histologic grade, eastern cancer oncology group, and plasma AFP levels. Furthermore, high E2F1 correlated with poor survival condition and pooled HR signified E2F1 as a risk factor for HCC. Enrichment analysis of differentially expressed genes, coexpressed genes, and putative targets of E2F1 emphasized the importance of cell cycle pathway, where CCNE1 and CCNA2 served as hub genes. Conclusions: We confirmed the upregulation of E2F1 and explored the prognostic value of E2F1 in HCC patients. Two putative targeted genes (CCNE1 and CCNA2) of E2F1 were identified for their potential roles in regulating cell cycle and promote antiapoptotic activity in HCC patients.

Background Immediate early response 3 (IER3) is correlated to the prognosis of several cancers, but the precise mechanisms underlying the regulation by IER3 of the occurrence and development of hepatocellular carcinoma (HCC) remain unknown. Methods The expression level of IER3 was examined by using in-house immunohistochemistry (IHC), public gene chip, and public RNA-sequencing (RNA-seq). The standardized mean difference (SMD) was calculated to compare the expression levels of IER3 between HCC patients and controls. The summary receiver operating characteristics (sROC) was plotted to comprehensively understand the discriminatory capability of IER3 between HCC and non-HCC group. The Kaplan–Meier curves and the combined hazard ratios (HRs) were used to determine the prognostic value of IER3 in HCC. Moreover, differentially expressed genes (DEGs) and co-expression genes (CEGs) were used to explored the molecular mechanisms of IER3 underlying HCC. hTFtarget was used to predict the transcription factors (TFs) of IER3. The binding site of TFs and the IER3 promoter region was forecasted using the JASPAR website. The relevant ChIP-seq data were used to determine whether TF peaks were present in the IER3 transcription initiation. Results A significantly increased expression of IER3 protein was found in HCC tissue relative to non-HCC tissue as detected by IHC (p < 0.001). Compared to 1,263 cases of non-HCC tissues, IER3 in 1483 cases of HCC tissues was upregulated (SMD = 0.42, 95% confidence interval [CI] [0.09–0.76]). The sROC showed that IER3 had a certain ability at differentiating HCC tissues (area under the curve (AUC) = 0.65, 95% CI [0.61–0.69]). Comprehensive analysis of the effect of IER3 on the prognosis of patients with HCC demonstrated that higher IER3 expression was associated with poor prognosis in HCC (HRs = 1.30, 95% CI [1.03–1.64]). Pathway enrichment analysis revealed that IER3-related genes were mostly enriched in the PI3K-Akt signaling pathway, cancer-related signaling pathways, the p53 signaling pathway, and other signaling pathways. Regulatory factor X5 (RFX5) was identified as a possible regulator of IER3-related TF. Conclusion IER3 may be a potential prognostic marker for HCC. The molecular mechanisms of IER3 in HCC warrant further study.

Abstract Background Ischemic cardiomyopathy (ICM) has ranked as the most common cause morbidity and mortality in the elderly over the past decades. One of the most important reasons for this is that its exact underlying mechanism remains poorly understood. Methods Five datasets were downloaded from the GEO database. Differential gene expression (DGE) was identified by the R RobustRankAggreg package. Differential miRNA expression was evaluated by the Limma package. Gene potential functions were then determined by the clusterProfiler database. The miRNA-DGE regulatory network was predicted by cyTargetLinker. Then, a protein-protein interaction network was constructed by STRING tool, MCODE, and BiNGO tool. Results 91 miRNAs and 274 potential genes were identified. Of these, COL1A1, IGF1 and CCND1 were found to be involved in many signaling pathways; and miR-9-5p was found to play critical roles in ICM. Conclusion Our study has unraveled the potential key genes and miRNAs as well as the possible underlying molecular pathogenesis of ICM, which is a crucial step leading to a new avenue for the early intervention of this disorder.

目的:探讨广西红水河流域长寿人群中HBB基因多态性对血清HBB mRNA表达水平的影响及其与代谢指标的关系.方法:采用实时荧光定量聚合酶链式反应(qRT-PCR)法对广西红水河流域长寿组168例及对照组196例进行血清HBB mRNA表达水平的检测,并使用PCR-Sanger测序法对HBB基因全长进行多态性检测,分析各多态性对HBB mRNA水平的影响及其与代谢指标的关系.结果:长寿组的血清HBB mRNA表达水平低于对照组(P＜0.05);HBB基因检测出6个多态,其中长寿组rs7480526A/A基因型的频率高于对照组(P＜0.05);长寿组rs33986703基因型T/T及等位基因T的频率低于对照组(均P＜0.05);长寿组rs7480526 A/A及rs33986703 T/T基因型的mRNA表达水平低于对照组(P＜0.05,P＜0.01);在总体人群中,rs7480526A/A基因型携带者的TG水平低于A/C携带者(P＜0.05),rs33986703T/T携带者的ApoA1水平高于T/A携带者(P＜0.05),这两个位点其他基因型携带者的代谢指标在长寿组及对照组之间无显著差异(P＞0.05).结论:HBB rs7480526及rs33986703基因多态可能通过表观遗传学影响HBB mRNA的表达,并参与对代谢指标的调控,进而影响衰老相关性疾病的发生发展及个体寿限.

BACKGROUND:The molecular mechanism of laryngeal squamous cell carcinoma (LSCC) is not completely clear, which leads to poor prognosis and treatment difficulties for LSCC patients. To date, no study has reported the exact expression level of zinc finger protein 71 (ZNF71) and its molecular mechanism in LSCC.METHODS:In-house immunohistochemistry (IHC) staining (33 LSCC samples and 29 non-LSCC samples) was utilized in analyzing the protein expression level of ZNF71 in LSCC. Gene chips and high-throughput sequencing data collected from multiple public resources (313 LSCC samples and 192 non-LSCC samples) were utilized in analyzing the exact mRNA expression level of ZNF71 in LSCC. Single-cell RNA sequencing (scRNA-seq) data was used to explore the expression status of ZNF71 in different LSCC subpopulations. Enrichment analysis of ZNF71, its positively and differentially co-expressed genes (PDCEGs), and its downstream target genes was employed to detect the potential molecular mechanism of ZNF71 in LSCC. Moreover, we conducted correlation analysis between ZNF71 expression and immune infiltration.RESULTS:ZNF71 was downregulated at the protein level (area under the curve [AUC] = 0.93, p < 0.0001) and the mRNA level (AUC = 0.71, p = 0.023) in LSCC tissues. Patients with nodal metastasis had lower protein expression level of ZNF71 than patients without nodal metastasis (p < 0.05), and male LSCC patients had lower mRNA expression level of ZNF71 than female LSCC patients (p < 0.01). ZNF71 was absent in different LSCC subpopulations, including cancer cells, plasma cells, and tumor-infiltrated immune cells, based on scRNA-seq analysis. Enrichment analysis showed that ZNF71 and its PDCEGs may influence the progression of LSCC by regulating downstream target genes of ZNF71. These downstream target genes of ZNF71 were mainly enriched in tight junctions. Moreover, downregulation of ZNF71 may influence the development and even therapy of LSCC by reducing immune infiltration.CONCLUSION:Downregulation of ZNF71 may promote the progression of LSCC by reducing tight junctions and immune infiltration; this requires further study.