Expression Levels In Tissues Research Articles

BTN1A1 expression in colon cancer: Implications for immunotherapy and patient stratification.

249 Background: Butyrophilin 1A1 (BTN1A1) is a promising immune checkpoint protein offering a novel option for immunotherapy. We discovered that BTN1A1 and PD-L1 expression are mutually exclusive through the JAK/STAT pathway in various tumors. Recently, Nelmastobart, an antibody targeting BTN1A1, successfully completed a Phase 1 clinical trial at three sites in the United States and two in the Republic of Korea, spanning 98 weeks. This abstract focuses on the results from the Phase 1 trial in patients with colorectal cancer (CRC), emphasizing biomarker diagnostics and therapeutics derived from translational research. Methods: We conducted a first-in-human, multicenter, open-label Phase I clinical trial to evaluate the safety, tolerability, pharmacokinetics, and preliminary efficacy of hSTC810 monotherapy in CRC patients. Extensive analyses of BTN1A1 expression were performed using patient specimens through immunohistochemistry and multi-immunofluorescence techniques to identify critical biomarkers and accurately assess nelmastobart's anticancer activity. We then analyzed progression-free survival (PFS), overall survival (OS), objective response rate (ORR), and safety alongside translational research findings using patient tissue samples. Results: A total of 47 patients were enrolled in the study. The median PFS across all cohorts was 1.9 months, while the median Kaplan-Meier estimate for OS was 12.8 months. Among the 16 CRC patients enrolled, tissue samples were obtained and analyzed using IHC in 12 patients and OPAL staining in 7 patients. The objective response rate (ORR) for CRC was 2.3%. The median progression-free survival (PFS) and overall survival (OS) were 3.6 months and 11 months, respectively. Notably, PFS, OS, and ORR strongly correlated with BTN1A1 expression levels in CRC patients. High levels of BTN1A1 expression in colorectal tumor tissues correlated with clinical outcomes, identifying BTN1A1 as a potential biomarker for predicting patient responses to targeted therapies. The mutual exclusivity of BTN1A1 and PD-L1 expression suggests that BTN1A1 could serve as a diagnostic marker to identify patients who may be unresponsive to PD-1/PD-L1 blockade therapy and might benefit from BTN1A1 blockade therapy. Conclusions: This Phase 1 clinical trial demonstrated that antibody-mediated BTN1A1 blockade effectively suppresses tumor growth in patients with BTN1A1-positive CRC. These findings suggest that BTN1A1-targeted therapies could provide a viable treatment option for patients with colon cancers that are refractory or resistant to anti-PD-1/PD-L1 therapies. Clinical trial information: NCT05231746 .

A phase I trial of intraperitoneal (IP) mesothelin (MSLN)-targeted CAR T-cell therapy in patients with MSLN-positive esophagogastric cancer (EGC) with peritoneal carcinomatosis.

TPS513 Background: Outcomes for patients with advanced EGC remain poor. The peritoneum is an immune-privileged niche and a common site of metastases occurring in up to 40% of patients with metastatic EGC; this is characterised by poor survival because of resistance to immunotherapy and a lack of measurable disease, which excludes trial participation. MSLN was selected as a target for CAR T cell therapy as it is overexpressed in up to 60% of EGC, is associated with aggressive disease biology and has low levels of expression in normal tissues. In pre-clinical in vivo models of peritoneal carcinomatosis, improved efficacy was observed with regional administration of IP CAR T cells, compared with systemic CAR T cells. No significant toxicity was observed at clinically relevant doses of IP administered CAR T-cells. M28z1XXPD1DNR CAR T-cells are a next-generation MSLN-targeted CAR, equipped with a modified CD3z (1XX), and a PD-1 dominant negative receptor (PD1DNR) that provides T-cell intrinsic checkpoint blockade. Methods: This is a single-arm phase I study assessing the safety, maximum tolerated dose (MTD) and preliminary efficacy of IP administered M28zXXPD1DNR CAR T cells in patients with MSLN-positive (IHC ≥ 25%) EGC with evidence of peritoneal carcinomatosis (imaging and/or cytology). Patients with concomitant extraperitoneal disease are eligible. Patients must have received at least one prior line of treatment for advanced or metastatic disease and have at least one measurable or evaluable lesion per RECIST 1.1. A minimum of 4 patients and maximum of 18 patients will be treated with 4 escalating doses (with 1 “fallback” dose) of CAR T cells to a maximum of 3 x 10 7 cells/kg administered via peritoneal catheter (Table). Lymphodepletion with fludarabine 30mg/m 2 /day and cyclophosphamide 300mg/m 2 /day is given for 3 days prior to the CAR T cell infusion. Adverse events, response rates (RECIST 1.1), survival outcomes and immuno-monitoring (immune cell phenotyping and serum/peritoneal fluid cytokine analysis) will be assessed. The trial is currently enrolling patients. Clinical trial information: Will be registered before ASCO GI. Dose-escalation scheme based on the continuous reassessment method. Dose Level Dose (cells/kg) 1 1 x 10 6 2 (Starting dose) 3 x 10 6 3 6 x 10 6 4 1 x 10 7 5 3 x 10 7

CARM1-induced lncRNA NEAT1 synchronously activates MYCN and GalNAcT-I to accelerate the progression of neuroblastoma.

Long non-coding RNAs (lncRNAs) play important roles in progression of neuroblastoma (NB). LncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) has been shown to affect the development of multiple tumors. However, the effect of NEAT1 on NB remain unclear. In this study, the new mechanisms whereby how NEAT1 influences tumor progression in NB was investigated. RT-qPCR, western blot, bioinformatics, cell growth, Transwell, and flow cytometric analyses were performed to determine how NEAT1 synchronously regulates the miR-873-5p/MYCN proto-oncogene(MYCN) and miR-873-5p/polypeptide N-acetylgalactosaminyltransferase 1(GalNAcT-I) axes to accelerate the progression of NB. NB-bearing animal models were established to evaluate the function of NEAT1 in NB. The relationships between transcription factor coactivatorassociated arginine methyltransferase 1 (CARM1) and NEAT1, NEAT1 and miR-873-5p, miR-873-5p and GalNacT-I or MYCN, were verified using luciferase reporter gene assay, respectively. Our study revealed elevated levels of NEAT1 expression in NB cells and tissues which was associated with an advanced pathological stage and poor prognostic outcomes. According to in vitro gain- and loss- of function experiments, NEAT1 enhances progression of NB. NEAT1 silencing was found to inhibit NB proliferation in vivo. Mechanistically, to achieve upstream regulation, epigenetic downregulation of NEAT1 was achieved via the inhibition of CARM1. NEAT1 was found to positively regulate MYCN and GalNAcT-I levels as a competitive sponge of miR-873-5p. Activity of the lncRNA NEAT1 can be triggered via CARM1, which synchronously promotes NB development via the miR-873-5p/MYCN and miR-873-5p/GalNAcT-I axes. These findings shed light on the novel molecular mechanisms underlying NB progression.

LncRNA RNF144A-AS1 gene polymorphisms and their influence on lung cancer patients in the Chinese Han Population

Lung cancer is primarily classified as NSCLC, which is distinguished by a wide range of genetic variations. This study focused on RNF144A-AS1, a relatively unexplored lncRNA, to explore the impact of its genetic polymorphisms on the susceptibility to NSCLC. We detected RNF144A-AS1 expression and its correlation with prognosis and clinical pathological features using bioinformatics analysis. The association between RNF144A-AS1 polymorphism and NSCLC susceptibility was evaluated using case-control methods. This investigation featured a cohort of 700 NSCLC individuals and 700 healthy controls. The genotype of genetic variation was detected by PCR-RFLP and iMLDR, followed by subsequent calculation of OR and 95 % CI. Our data show that RNF144A-AS1 exhibits high expression levels in LUAD tissues and its expression is closely linked to LUAD progression and prognosis. Carrier of RNF144A-AS1 rs3806609 TT genotype increased NSCLC susceptibility compared to carrier of rs3806609 CC genotype (OR=2.21, 95%CI=1.57-3.13). Our study identifies RNF144A-AS1 genetic variants as potential susceptibility markers in NSCLC. RNF144A-AS1 promotes cell proliferation and migration in LUAD through the IFN-γ/JAK2/STAT1 signalling pathway. Collectively, these findings pave the way for developing targeted therapies and diagnostic tools based on RNF144A-AS1 and its variants.

Identification and functional analysis of a novel L-type lectin (NdLTL1) from Neocaridina denticulata sinensis.

This study investigates an L-type lectin, NdLTL1, derived from Neocaridina denticulata sinensis, emphasizing its role in immune defense through carbohydrate binding and bacterial agglutination. Bioinformatics analysis identified 179 lectin sequences, leading to subsequent investigations into the structure and function of NdLTL1. The open reading frame (ORF) of NdLTL1 spans 966 bp and encodes a protein consisting of 321amino acids (36.25kDa), which features a signal peptide, a transmembrane domain and Lectin_leg-like domain. Three-dimensional modeling revealed three antiparallel β-sheets characteristic of Lectin_leg-like domain, confirming evolutionary links with proteins such as VIP36. Protein-carbohydrate and protein-protein interaction studies showed that NdLTL1 binds to both carbohydrates like N-acetylglucosamine, peptidoglycan, lipopolysaccharides (LPS), and mannose, as well as sorting proteins (COPI/COPII). Gene expression analyses indicated that NdLTL1 exhibits the highest expression levels in cardiac tissues and significant upregulation in gills following exposure to Vibrio parahaemolyticus. Recombinant NdLTL1 expressed in Escherichia coli was shown to bind multiple bacterial strains and exhibit calcium-dependent agglutination properties. Enzyme-linked immunosorbent assay (ELISA) confirmed concentration-dependent carbohydrate binding, particularly rapid for LPS. In vitro experiments suggested that recombinant NdLTL1 may promote bacterial growth under nutrient-limited conditions while potentially triggering immune defenses indirectly.

Distinctive Lipogenic Gene Expression Patterns in the Mammary Glands of Dairy Cows Are Associated with the Unique Fatty Acid Composition of Bovine Milk Fat

Fat composition is largely responsible for the technological and rheological properties of cow milk and dairy products. Bovine milk fat is unique in terms of its fatty acid composition and positional distribution, with about 25% of its fatty acids being short- and medium-chain, which are synthesized de novo in the mammary gland and are not present in extra-mammary tissues. With the aim to identify potential genetic factors responsible for the unique composition of bovine milk fat, we extracted genes with GO annotations related to lipid metabolism and performed a gene expression mega-analysis. Overall, different lipogenic tissues (i.e., mammary, liver, and adipose) displayed discerned expression patterns. In a PCA, the liver was significantly separated from adipose and mammary tissues. In a correlation analysis with the fatty acid synthetase (FASN) gene, notable differences among the tissues were found. In the mammary gland, the majority of genes (~70%) were negatively correlated with FASN expression, whereas only 18% were negatively correlated in adipose. Only a few genes were positively correlated with FASN exclusively in the mammary gland, including AGPAT1 and AGPAT6, which also had the highest expression in the mammary gland compared with adipose. Looking at the expression levels in tissues (TPM) revealed significant differences in the expressions of genes responsible for the activation of fatty acids by ligation to CoA, according to their carbon chain length. Notably, the ACSS1 gene, which converts acetate to acetyl-CoA, had the highest expression in the mammary gland, whereas genes responsible for the activation of long-chain fatty acids had lower expressions. The findings of the present study suggest that the unique properties of dairy fat are the results of the distinct expression patterns of genes involved in de novo synthesis of fatty acids and their downstream utilization.

The Clinical Diagnostic Value of the Super-enhancer-associated Long Noncoding RNA RP11-803D5.4 and AC005592.2 in Colorectal Cancer.

Super-enhancer-associated long noncoding RNAs (SE-lncRNAs) play crucial roles in CRC pathogenesis. RP11-803D5.4 and AC005592.2 were identified as SE-lncRNAs of interest via microarray analysis, and our study aimed to evaluate their clinical value in CRC diagnosis and prognosis assessment. Fluorescence quantitative real-time PCR (qRT-PCR) was used to measure the expression of RP11-803D5.4 and AC005592.2 in the tissues and serum of CRC patients. Receiver operating characteristic (ROC) curves were generated to determine the predictive value of the two SE-lncRNAs. Functional assays were applied to assess the ability of RP11-803D5.4 to promote the proliferation, migration, and invasion of CRC cells. The two SE-lncRNAs were significantly upregulated in CRC tissue and serum samples vs. corresponding controls. ROC curve analysis indicated that RP11-803D5.4 (AUC=0.842) and AC005592.2 (AUC=0.811) had a high diagnostic performance for CRC. The combination of RP11-803D5.4, AC005592.2, and CEA had an AUC of 0.946 and distinguished CRC patients and healthy controls better than SE-lncRNA alone. The serum levels of RP11-803D5.4 and AC005592.2 were strongly correlated with their tissue expression levels. The expression levels of the two SE-lncRNAs were significantly lower in postoperative samples than in preoperative samples. Furthermore, similar to the findings of previous studies on AC005592.2, high RP11-803D5.4 expression promoted the proliferation, invasion, and migration of CRC cells. The findings suggested that RP11-803D5.4 and AC005592.2 are upregulated in CRCact and are crucial promoters of CRC progression. They also suggested that they might serve as noninvasive biomarkers for diagnosing CRC.

Integrating E-cadherin expression levels with TNM staging for enhanced prognostic prediction in colorectal cancer patients

PurposeThis study aimed to investigate the clinical significance of E-cadherin expression levels in colorectal cancer tissues and explore the relationship between E-cadherin expression and tumor node metastasis (TNM) stage. The goal was to establish a more accurate prognostic prediction for colorectal cancer patients by analyzing E-cadherin expression levels alongside TNM staging.MethodsThe study examined colorectal cancer patients by dividing them into groups based on E-cadherin expression levels. It then assessed their 5-year event-free survival (EFS) and disease-specific survival (DSS) hazard ratios (HRs). Additionally, the prognosis of patients was analyzed by combining E-cadherin expression levels with TNM staging, particularly focusing on patients in stages III and IV.ResultsThe E-cadherinLow group had significantly worse outcomes, with HRs of 2.30 for EFS and 2.76 for DSS compared to the E-cadherinHigh group. When combined with TNM stage III/IV, patients with E-cadherinLow expression showed a poor prognosis, with HRs of 1.93 for EFS and 2.35 for DSS compared to those with E-cadherinHigh expression at the same TNM stage.ConclusionsLow levels of E-cadherin expression are associated with a poor prognosis and decreased survival in colorectal cancer patients. Combining E-cadherin expression levels with TNM staging provides a more precise prediction of patient prognosis and survival, potentially guiding personalized treatment strategies.

A whole-body imaging technique for tumor-specific diagnostics and screening of B7-H3-targeted therapies.

B7-H3 or CD276 is notably overexpressed in various malignant tumor cells in humans, with extremely high expression rates. The development of a radiotracer that targets B7-H3 may provide a universal tumor-specific imaging agent and allow the noninvasive assessment of the whole-body distribution of B7-H3-expressing lesions. We enhanced and optimized the structure of an affibody (ABY) that targets B7-H3 to create the radiolabeled radiotracer [68Ga]Ga-B7H3-BCH, and then, we conducted both foundational experiments and clinical translational studies. [68Ga]Ga-B7H3-BCH exhibited high affinity (Kd = 4.5 nM), and it was taken up in large amounts by B7-H3-transfected cells (A549CD276 and H1975CD276 cells); these phenomena were inhibited by unlabeled precursors. Moreover, PET imaging of multiple xenograft models revealed extensive [68Ga]Ga-B7H3-BCH uptake by tumors. In a clinical study including 20 patients with malignant tumors, the [68Ga]Ga-B7H3-BCH signal aggregated in both primary and metastatic lesions, surpassing 18F-FDG in overall diagnostic efficacy for tumors (85.0% vs 81.7%), including differentiated hepatocellular and metastatic gastric cancers. A strong correlation between B7-H3 expression and [68Ga]Ga-B7H3-BCH uptake in tumors was observed, and B7-H3 expression was detected with 84.38% sensitivity and 100% specificity when an SUVmax of 3.85 was set as the cutoff value. Additionally, B7-H3-specific PET imaging is expected to predict B7H3 expression levels in tumor cells, intratumoral stroma and peritumoral tissues. In summary, [68Ga]Ga-B7H3-BCH has potential for the noninvasive identification of B7H3 expression in systemic lesions in patients with malignant tumors. This agent has prospects for improving pretreatment evaluation, predicting therapeutic responses, and monitoring resistance to therapy in patients with malignancies. gov NCT06454955. This research was financially supported by the Natural Science Foundation of Beijing Municipality (No. 7242266), the National Natural Science Foundation of China (No. 82202201), and the Young Elite Scientists Sponsorship Program by CAST (No. YESS20220230).

Effect of selenium on the dysfunction of rat salivary glands induced by 131I and expression of insulin-like growth factors and aquaporins.

To investigate the effects of selenium on functional and histopathological changes and mRNA expression levels of insulin-like growth factors 1 and 2 (IGF-1 and -2) and aquaporins 4 and 5 (AQP-4 and -5) in 131I-induced damaged rat parotid glands. Rats were divided into three groups: iodotherapy-with-selenium, iodotherapy-only, and control. Rats in the iodotherapy-with-selenium group were intragastrically administered 131I on the first day and selenomethionine through drinking water. Rats in the iodotherapy-only group were only administered 131I. Changes in parotid gland function were evaluated using the functional parameters of salivary gland dynamics imaging pre-experiment and on days 7, 30, and 90 post-treatment. Immunofluorescence and quantitative real-time PCR analyses detected IGF-1, IGF-2, AQP-4, and AQP-5 expression levels in tissues. The gland-to background ratio at a maximum count (G/BGmax), Tmax/Tmin, and Smax values were significantly impacted over time in the iodotherapy-with-selenium group; on day 30, the G/BGmax value was significantly higher than that in the iodotherapy-only group. Histopathological analysis revealed that on days 30 and 90, the iodotherapy-with-selenium group displayed greater parotid gland repair than the iodotherapy-only group. In the iodotherapy-with-selenium group, fluorescence intensity and mRNA levels of AQP-5 increased with the selenium supplementation period, reaching significantly higher levels on days 30 and 90 than in the iodotherapy-only group. Whereas the fluorescence intensity and mRNA levels of IGF-1 in the iodotherapy-with-selenium group were significantly higher on day 7 than on day 30 in the iodotherapy-only group. Selenium may repair 131I-induced tissue and functional damage in rat salivary glands by upregulating AQP-5 and IGF-1 expression.

Genome-wide analysis of the SPL family in Zanthoxylum armatum and ZaSPL21 promotes flowering and improves salt tolerance in transgenic Nicotiana benthamiana.

Z. armatum is an economically valued crop known for its rich aroma and medicinal properties. This study identified 45 members of the SQUAMOSA-PROMOTER BINDING PROTEIN LIKE (SPL) gene family in the genome of Z. armatum. Phylogenetic and collinearity analyzes demonstrated a close relationship between ZaSPLs and ZbSPLs from B subgenomes of Zanthoxylum bungeanum. Our miRNA sequencing revealed a high degree of conservation of miR156a within Z. armatum, with the za-miR156a sequence identical to miR156-5p in Arabidopsis thaliana and Citrus sinensis. Of the 45 genes identified by ZaSPLs, 21 were targeted by za-miR156a, transient co-expression experiments in N. benthamiana demonstrated the targeting relationship between za-miR156 and ZaSPL21. Furthermore, RNA-seq and qRT-PCR analysis revealed that ZaSPL genes exhibited elevated expression levels in juvenile tissues of Z. armatum. The expression of nine representative ZaSPL genes were upregulated under polyethylene glycol (PEG) and abscisic acid (ABA). Overexpression of ZaSPL21 delayed the germination of transgenic tobacco and facilitated the flowering process in transgenic N. benthamiana. Significant up-regulation in the expression levels of flowering-related genes such as NbFT1, NbPIP2;1, NbTCP1, NbCOL1, NbGI2, NbGAI1, NbCKX2, and NbARR4 was observed in transgenic plants, suggesting that ZaSPL21 may stimulate plant flowering by regulation of these genes. Furthermore, ZaSPL21 also increased the germination speed of transgenic tobacco seeds during drought and salt stress conditions, and improved the salt tolerance of transgenic seedlings. In conclusion, our study contributes to understanding the functional analysis of the SPL gene family in Z. armatum and emphasizes the crucial role of ZaSPL21 in improving tolerance to salt and promoting flowering. The results offer potential strategies for the further utilization of these genes to improve the salt tolerance of Z. armatum.

Overexpression of CHMP2B suppresses proliferation of renal clear cell carcinoma cells.

To analyze the association of CHMP2B expression level of with clinicopathological characteristics and prognosis of clear cell renal cell carcinoma (CRCC) and the possible role of CHMP2B in tumorigenesis and progression of CRCC. RNAseq data of CRCC were downloaded from the TCGA database for analysis of CHMP2B expression levels in tumor and adjacent tissues and their correlation with clinicopathological characteristics of the patients. Survival outcomes of the patients with high and low CHMP2B expressions were analyzed using the Kaplan-Meier model, and the COX risk regression model was used for identifying the prognostic factors of the patients. The correlation between CHMP2B expression and immune infiltration, its co-expressed genes, and the effect of CHMP2B gene mutations on immunotherapy responses, and its immunohistochemical expression in CRCC and normal tissues were analyzed. Clinical samples of CRCC were collected to examine CHMP2B expressions using RT-PCR, and cell experiment was carried out to test the effect of CHMP2B overexpression on biological behaviors of CRCC cells. CHMP2B was significantly under-expressed in renal cancer tissues, and its overexpression obviously inhibited the proliferation of CRCC cells in vitro. CHMP2B expression level was significantly correlated with age, gender, lymph node metastasis, and tumor stage, and the patients with low CHMP2B expression had poor survival outcomes. Enrichment and co-expression gene analyses suggested that CHMP2B was mainly involved in viral outgrowth, necrotic apoptosis, endocytosis, and immune-regulatory processes in kidney cancer. CHMP2B is lowly expressed in renal cancer tissues to affect tumor progression and tumor immune processes, and may serve as a prognostic biomarker and therapeutic target for CRCC.

M6A-modified lncRNA GAS5 promotes M1-polarization of microglia in alcohol use disorder.

Long noncoding RNA (lncRNA) are essential for modulating the onset and progression of alcohol use disorder (AUD). In this study, we investigated the molecular pathways through which lncRNA may contribute to AUD development. We assessed the expression levels of long noncoding RNA GAS5 (lncRNA GAS5) and microRNA-136-5p (miR-136-5p) in AUD tissue samples and cell lines using reverse transcription-quantitative polymerase chain reaction. Detection of GAS5 N6-methyladenosine (m6A) modifications, facilitated by alkylation repair homolog 5 (ALKBH5), was performed using RNA immunoprecipitation and RNA pull-down assays. The effect of GAS5 on the functionality of SH-SY5Y cells was evaluated using CCK-8 and Transwell assays. Our findings showed high levels of GAS5 expression in both AUD tissues and cell lines. Overexpression of GAS5 decreased the migratory capability of SH-SY5Y cells, whereas silencing GAS5 increased this ability. Bioinformatics analyses predicted a relationship between expression levels of miR-136-5p and GAS5, which was subsequently confirmed using dual-luciferase reporter assays. Additionally, we discovered that GAS5 acts as a sponge for miR-136-5p, leading to the upregulation of ATF2. Elevated levels of ATF2 are associated with M1 microglial polarization. In summary, m6A-modified GAS5 may influence the M1 polarization of microglia via the miR-136-5p/ATF2 pathway. Statistical evaluations were performed using GraphPad Prism V8.0, employing the student's t-test for comparisons between two groups, assuming a normal distribution and equal variances. When variances were unequal, but normality was maintained, the corrected Student's t-test was applied. The non-parametric Wilcoxon rank-sum test was used to analyze non-normally distributed data, and one-way ANOVA was used to compare three or more groups. Independent replication was ensured in the studies, with each experiment repeated at least three times and statistical significance was set at P < 0.05.

Comprehensive investigation of matrix metalloproteinases in skin cutaneous melanoma: diagnostic, prognostic, and therapeutic insights

The dysregulation of matrix metalloproteinases (MMPs) in skin cutaneous melanoma (SKCM) represents a critical aspect of tumorigenesis. In this study, we investigated the diagnostic, prognostic, and therapeutic aspects of the MMPs in SKCM. Thirteen SKCM cell lines and seven normal skin cell lines were cultured under standard conditions for experimental analyses. RNA and DNA were extracted, followed by RT-qPCR to assess MMP expression and promoter methylation analysis to determine methylation levels. Functional assays, including cell proliferation, colony formation, and wound healing, were conducted post-MMP7 knockdown using siRNA in A375 cells. Databases like GEPIA2, HPA, MEXPRESS, and miRNet were employed for expression, survival, methylation, and miRNA-mRNA network analyses. We investigated the expression and promoter methylation landscape of MMPs in SKCM cell lines, revealing significant (p-value < 0.05) up-regulation of MMP1, MMP7, MMP9, MMP10, MMP11, MMP12, MMP13, MMP14, and MMP25, alongside down-regulation of MMP2, MMP3, and MMP21. Furthermore, our analysis demonstrated a significant (p-value < 0.05) inverse correlation between MMP expression levels and promoter methylation status, suggesting a potential regulatory role of DNA methylation in MMP dysregulation. Notably, MMP7, MMP11, and MMP14 exhibited significant (p-value < 0.05) associations with the overall survival of SKCM patients, emphasizing their prognostic significance. Additionally, Receiver operating characteristic (ROC) curve analysis highlighted the significant (p-value < 0.05) diagnostic potential of MMP7, MMP11, and MMP14 in distinguishing SKCM from normal individuals. Subsequent validation across multiple cohorts confirmed significant (p-value < 0.05) elevated MMP expression levels in SKCM tissues, particularly in advanced disease stages, further emphasizing their role in tumor progression. Furthermore, we elucidated potential regulatory pathways involving miR-22-3p, which targets MMP7, MMP11, and MMP14 genes in SKCM. Our findings also revealed associations between MMP expression and immune modulation, drug sensitivity, and functional states of SKCM cells. Lastly, MMP7 knockdown in A375 cells significantly significant (p-value < 0.05) impacted several characteristics, including cell proliferation, colony formation, and wound healing. Our findings highlight the diagnostic, prognostic, and therapeutic potential of MMP7, MMP11, and MMP14 in SKCM. These MMPs could serve as biomarkers for early detection and targets for therapy. Future efforts should focus on preclinical and clinical validation to translate these insights into personalized diagnostic and therapeutic strategies.

Expression level of miR-548aa in tissue samples of patients with colorectal cancer.

The pathogenesis of colorectal cancer (CRC) is influenced by various risk factors, and genetic alterations in progression of colon polyps. The expression patterns of microRNA-548 (miR-548) in colorectal tissues have been sufficiently characterized. The aim of this study is to clarify the role of miR-548aa in tumorigenesis, gene targeting, predictive value and its expression levels in tumoral versus adjacent marginal tissues in CRC patients. This study included 35 CRC patients who underwent surgery to remove their tumor tissue. Tumor samples and adjacent marginal tissue were collected and gene expression was analyzed through real-time PCR. The correlation between miR-548aa expression levels and clinical parameters were investigated. Our findings showed a significant increase in the expression of miR-548aa in tumoral tissues compared to marginal tissues (p < 0.05). The upregulation of miR-548aa was significantly detected in CRC samples, showing an area under the curve of 0.89 (p = 0.002), indicating strong sensitivity and specificity for CRC diagnosis. To prediction of miRNA target genes and construction of regulatory networks of miR-548aa, we used bioinformatics tools including TargetScan and miRDB. Protein-protein interaction (PPI) network was constructed through STRING database and visualized using Cytoscape software. Dysregulation of miR-548aa is closely related to tumorigenesis in colorectal tissues, which affects disease progression and clinical outcomes. The miR-548aa can be introduced as a proposed biomolecule involved in mechanism of tumorigenesis, gene targeting and predictive value for CRC diagnosis and a target for therapeutic intervention.

CircCCT2/miR-146a-5p/IRAK1 axis promotes the development of head and neck squamous cell carcinoma

BackgroundHead and neck squamous cell carcinoma (HNSCC), a highly invasive malignancy with a poor prognosis, is one of the most common cancers globally. Circular RNAs (circRNAs) have become key regulators of human malignancies, but further studies are necessary to fully understand their functions and possible causes in HNSCC.MethodsCircCCT2 expression levels in HNSCC tissues and cells were measured via qPCR. CircCCT2 was characterized by Sanger sequencing, qRT-PCR, RNase R & Actinomycin D treatment, nucleoplasmic separation and FISH experiments. CCK-8 and colony formation assays were performed to determine cell proliferation, and Transwell assays were used to determine migration and invasion. A xenograft tumor model was used to study the influence of circCCT2 on HNSCC in vivo. Dual-luciferase gene reporter, RIP, western blotting, and rescue experiments, were used to explore target-binding relationships and regulatory mechanisms.ResultsCircCCT2 was significantly upregulated in HNSCC tissues and cells. High circCCT2 levels were associated with advanced T stage, N stage, clinical stage and poor prognosis. Functionally, we verified that circCCT2 promotes HNSCC development in vitro and in vivo. Mechanistically, functioning as a competitive endogenous RNA (ceRNA) or miRNA sponge, circCCT2 binds directly to miR-146a-5p and increases interleukin-1 receptor-associated kinase 1 (IRAK1) levels, which enhances the malignant development of HNSCC by driving epithelial-mesenchymal transition (EMT).ConclusionCircCCT2 promotes HNSCC development through the miR-146a-5p/IRAK1 axis, revealing that circCCT2 is a potential biomarker and target for HNSCC.

LncRNA MANCR is downregulated in non-small cell lung cancer and predicts poor survival

BackgroundIt is known that genomic instability contributes to cancer development. Mitotically associated long non-coding RNA (MANCR) has been reported to promote genomic stability, suggesting its involvement in cancers. Therefore, this study was conducted to investigate the role of MANCR in non-small cell lung cancer (NSCLC).MethodsAfter NSCLC (n = 60) and control (healthy subjects, n = 60) plasma samples, as well as NSCLC and paired non-tumor tissues from patients were collected, the levels of MANCR expression in plasma and tissues was detected using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Then the correlations of MANCR expression with clinical stages were confirmed. The diagnostic values of MANCR in both plasma and tissue samples for stage I/II NSCLC were analyzed using Receiver Operating Characteristic (ROC) curves. All NSCLC patients were monitored for 5 years to investigate the role of MANCR in the prediction of patients’ survival.ResultsMANCR expression was downregulated in both NSCLC plasma and tissue of NSCLC patients compared to controls (P < 0.05). Decreased MANCR expression levels from stage I to IV were observed. However, MANCR expression in non-tumor tissue was not significantly different between different stages (P > 0.05). Additionally, stage I/II NSCLC patients were separated from controls using MANCR in plasma and tumor tissues as biomarkers. Lower MANCR levels in plasma and tumor were closely correlated with patients’ higher mortality rate.ConclusionMANCR is down-expressed in NSCLC patients and may serve as a diagnostic and prognostic biomarker for NSCLC.

Investigation of the Mesencephalic Astrocyte Derived Neurotrophic Factor-Endoplasmic Reticulum Stress Pathway in Mood Disorders.

Bipolar disorder (BD) has been associated with impaired cellular resilience. Recent studies have shown abnormalities in the unfolded protein response (UPR) in BD. The UPR is the cellular response to endoplasmic reticulum (ER) stress. Mesencephalic astrocyte-derived neurotrophic factor (MANF), a trophic factor, decreases ER stress by modulating the UPR. The objective of this study is to investigate the MANF-ER stress pathway in BD and major depressive disorder (MDD) compared to healthy controls (HC). MANF protein concentration and MANF and GRP78 gene expression were assessed in peripheral blood from individuals with BD, MDD and HC (protein: 40 BD, 55 MDD, 55 HC; gene expression: 52 BD, 61 MDD, 69 HC). MANF protein and gene expression along with GRP78 gene expression were also analyzed in postmortem brain tissue (20 BD, 20 MDD, 19 HC). MANF protein was quantified using an ELISA assay while quantitative polymerase chain reaction was used for MANF and GRP78 gene expression. Peripheral MANF protein levels were reduced in individuals with BD in a depressive state compared to controls (p=0.031) and euthymic BD participants (p=0.013). No significant differences in MANF or GRP78 gene expression were observed in BD irrespective of mood state, or MDD compared to HC (all p>0.05). No differences were observed regarding MANF/GRP78 protein or gene expression levels in postmortem tissue (p>0.05). Individuals with BD who were in an acute depressive phase were found to have reduced peripheral MANF levels potentially signifying abnormal UPR and supporting the notion that BD is associated with increased ER stress.

Resveratrol alleviates blast lung injury by modulating the epithelial sodium channel (ENaC) via the PI3K/AKT pathway.

Blast lung injury (BLI) is a major cause of death in blast injuries, largely due to pulmonary edema. Effective clearance of alveolar fluid is critical for resolving pulmonary edema, with the epithelial sodium channel (ENaC) playing a key role in this process. Resveratrol (RES), a natural compound with known antioxidant and anti-inflammatory properties, has shown promise in treating respiratory diseases. However, its potential protective effects against BLI, particularly through ENaC modulation, remain unclear. In this study, complementary in vivo and in vitro models were used to investigate mechanisms underlying pulmonary edema following a gas explosion. We discovered that RES significantly alleviated BLI by decreasing pulmonary edema, pulmonary index, W/D ratio, oxidative stress, inflammatory cell infiltration, and TNF-α and IL-1β expression levels in rat lung tissue. RES upregulated the expression of ENaC and PI3K/AKT both in rats and in A549 cells. The knockout of the PI3K-p85 gene suppressed RES-induced upregulation of p-AKT and ENaC, reversing protein expression in A549 cells. Altogether, RES shows potential to attenuate blast-induced lung injury by upregulating ENaC, partly through the PI3K/AKT signaling pathway. This study highlights RES as a potential therapeutic agent for BLI and offers new insights into its mechanisms of action.

Hypoxic BMSC-derived exosomal miR-210-3p promotes progression of triple-negative breast cancer cells via NFIX-Wnt/β-catenin signaling axis

BackgroundBone marrow mesenchymal stem cells (BMSCs) are a crucial component of the tumor microenvironment (TME), with hypoxic conditions promoting their migration to tumors. Exosomes play a vital role in cell-to-cell communication within the TME. Hypoxic TME have a great impact on the release, uptake and biofunctions of exosomes. This study aims to elucidate the communication between BMSC-derived exosomal miRNA and triple-negative breast cancer (TNBC) in a hypoxic environment.MethodsExosomes were isolated via ultracentrifugation and identified using scanning electron microscopy (SEM), nanoparticle tracking analysis (NTA) and western blot. A range of bioinformatics approaches were used to screen exosomal miRNAs and the target mRNAs of miRNAs and predict the possible signaling pathways. Expression levels of genes and proteins were assessed by quantitative real-time PCR and western blot. Cell proliferation, apoptosis, migration and invasion were analyzed using CCK-8 assay, EDU assay, transwell migration, wound healing assay and invasion assay, respectively. Dual luciferase reporter gene assay was conducted to confirm the binding between miRNAs and the target mRNAs. The impact of hypoxic BMSC-derived exosomal miRNA on TNBC progression in vivo was evaluated using tumor xenograft nude mouse models. Furthermore, the impact of patients’ serum exosomal miRNA on TNBC was implemented.ResultsExosomes derived from hypoxic BMSCs promotes the proliferation, migration, invasion and epithelial-mesenchymal transition of TNBC and suppresses the apoptosis of TNBC. The expression of miR-210-3p in BMSC-derived exosomes is markedly elevated in hypoxic conditions. Exosome-mediated transfer of miR-210-3p from hypoxic BMSCs to TNBC targets NFIX and activates Wnt/β-Catenin signaling in TNBC. Deletion of miR-210-3p in hypoxic BMSC-derived exosomes attenuates TNBC in vivo. Additionally, human exosomal miR-210-3p from the serum of TNBC patients promotes TNBC progression. Moreover, we notably observed a marked downregulation of NFIX expression levels in cancerous tissues compared to paracancerous tissues.ConclusionsHypoxic BMSC-derived exosomal miR-210-3p promotes TNBC progression via NFIX-Wnt/β-catenin signaling axis.