Analysis Of Protein Expression Research Articles

WDR62 mediates MAPK/ERK pathway to stimulate DNA damage repair and attenuate cisplatin sensitivity in lung adenocarcinoma.

Chemotherapy resistance has long stood in the way of therapeutic advancement for lung cancer patients, the malignant tumor with the highest incidence and fatality rate in the world. Patients with lung adenocarcinoma (LUAD) now have a dismal prognosis due to the development of cisplatin (DDP) resistance, forcing them to use more costly second-line therapies. Therefore, overcoming resistance and enhancing patient outcomes can be achieved by comprehending the regulatory mechanisms of DDP resistance in LUAD. WD repeat domain 62 (WDR62) expression in LUAD tissues and in DDP-resistant or sensitive LUAD patients was analyzed bioinformatically, and a K-M plot was utilized to assess survival status. Real-time quantitative PCR was employed for WDR62 expression detection, cell-counting kit-8 assay for half maximal inhibitory concentration determination, flow cytometry for cell apoptosis detection, immunofluorescence for γ-H2AX expression analysis, and western blot for nonhomologous end joining repair and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway-related protein expression analysis. Poor prognosis was linked to WDR62, which was overexpressed in LUAD tissues and cells. Compared to sensitive cells, DDP-resistant cells had increased WDR62 expression. WDR62 knockdown may enhance DDP-induced cell apoptosis while reducing cell proliferation and DNA damage repair. Functional investigations verified that overexpressed WDR62's encouraging impact on DNA damage repair in A549/DDP cells could be reversed by MAPK inhibitors, increasing the cells' susceptibility to DDP. LUAD cells became less sensitive to DDP when WDR62 activated the MAPK/ERK pathway, which promoted DNA damage repair, indicating that DDP resistance might be reversed by treating LUAD with inhibitors of the MAPK pathway.

Long-range directional growth of neurites induced by magnetic forces.

The ability to control the growth and orientation of neurites over long distances has significant implications for regenerative therapies and the development of physiologically relevant brain tissue models. In this study, the forces generated on magnetic nanoparticles internalised within intracellular endosomes are used to direct the orientation of neuronal outgrowth in cell cultures. Following differentiation, neurite orientation was observed after 3 days application of magnetic forces to human neuroblastoma (SH-SY5Y) cells, and after 4 days application to rat cortical primary neurons. The direction of neurite outgrowth was quantified using a 2D Fourier transform analysis, showing agreement with the derived magnetic force vectors. Orientation control was found to be effective over areas >1cm2 using modest forces of ∼10 fN per endosome, apparently limited only by the local confluence of the cells. A bioinformatics analysis of protein expression in cells exposed to magnetic forces revealed changes to cell signaling and metabolic pathways resulting in enhanced carbohydrate metabolism, as well as the perturbation of processes related to cellular organisation and proliferation. Additionally, in cell culture regions where the measured force vectors converged, large (∼100 µm) SH-SY5Y neuroclusters loaded with nanoparticles were found, connected by unusually thick linear neurite fibres. This could suggest a magnetically driven enhancement of neurocluster growth, with the clusters themselves contributing to the local forces that direct outgrowth. Such structures, which have not been previously observed, could provide new insights into the development and possible enhancement of neural circuitry. STATEMENT OF SIGNIFICANCE: A magnetic force approach for directing outgrowth in neuronal cells over macroscopic areas is successfully demonstrated. Cells were incubated with magnetic nanoparticles which were sequestered into intracellular compartments. Permanent magnet arrays created local intracellular magnetic force vectors mediated via the internalized nanoparticles, which were found to precisely guide neurite orientation. Analysis of cellular protein expression suggested the mechanism for directed growth involved specific cell signaling and metabolic pathways. In addition, highly unusual straight and thick neural fibers were observed that connected large 'magnetic' spherical cell clusters. The results reported will advance nanotechnology and cell therapy for neuro-regeneration where magnetic forces could help to reconnect damaged neurons, or even build artificial neuronal architectures.

Isolation of Endothelial Cells from Human Internal Mammary Artery.

Endothelial cells are a major contributor to cardiovascular diseases. Studying their function and how they influence pathological processes remains an ongoing area of research. Although primary endothelial cells might be readily available from animals, translating results from these studies can be challenging due to species-dependent differences. A common source of human endothelial cells is human umbilical vein endothelial cells, but these cells might not represent the variety of endothelial cells from various vascular beds. Here we describe a protocol for the isolation and cultivation of endothelial cells from human internal mammary artery remains. These remains are commonly available from coronary artery bypass graft surgeries. The described tissue explant method combined with a magnetic sorting process provided a relatively high yield of endothelial cells that were subsequently passaged and used for studies involving mRNA and protein expression analyses, as well as fluorescence-based immunohistochemistry and patch-clamp electrophysiology. This protocol may help to extend the repertoire of studying human endothelial cells to understand their role and function in health and disease.

Comprehensive drug-like assessment of pyridine carbothioamide analogs: from molecular modeling to in-vivo evaluation.

To evaluate the anti-inflammatory potential of novel class of chemical compounds designed by the linkage of carbothioamide moiety with pyridine. In silico analysis was conducted using molecular docking followed by an in vitro cytotoxicity assay and evaluation of anti-inflammatory activity. Subsequently, in vivo performance was determined using the Complete Freund's Adjuvant-induced inflammatory model, employing macroscopic, histopathological, and protein expression analyses. Molecular interaction studies revealed that compound R2 displayed the most favorable binding mode with human nitric oxide synthase, cyclooxygenase-1, and cycloxygenase-2. All compounds exhibit dose-dependent cytotoxicity. Notably, compound R4 was safer at higher concentration, whereas compound R2 was comparatively toxic. The in vitro anti-inflammatory activity demonstrated half maximal inhibitory concentration (IC50) values ranging from 10.25 ± 0.0 to 23.15 ± 4.24 µM, with compound R6 exhibiting the lowest IC50 value and compound R3 showing the highest. The in vivo results corroborated the anti-inflammatory effects, with a significant reduction in paw size (p < 0.001). Among the tested compounds, compound R4 exhibited the most potent anti-inflammatory activity, whereas R2 exhibited the least potency. The study highlights the promise of discovering new anti-inflammatory drugs containing pyridine moiety with proven potency, efficacy, and reduced side effects.

Glycosylation Patterns in Meccus (Triatoma) pallidipennis Gut: Implications for the Development of Vector Control Strategies

The primary mode of transmission for Chagas disease is vector-borne transmission, spread by hematophagous insects of the Triatominae subfamily. In Mexico, the triatomine Meccus pallidipennis is particularly significant in the transmission of Trypanosoma cruzi. This study focused on analyzing protein expression and modifications by glycosylation in different regions of the digestive tract of fifth-instar nymphs of M. pallidipennis. Two gut sections were dissected and extracted: the anterior midgut (AMG) and the proctodeum or rectum (RE). Proteins were extracted from each tissue sample and profiled by one- and two-dimensional electrophoresis; protein glycosylation was analyzed by lectin affinity. Our results showed significant differences in protein expression and glycosylation between both gut regions, with modifications being more frequent in the RE. The proteins HSP70, actin, and tubulin were analyzed, finding a differential expression of the latter two between AMG and RE. Understanding glycosylation patterns provides critical insights into vector–pathogen interactions that could eventually inform novel control approaches. Furthermore, the potential use of lectins as insecticidal agents highlights the broader implications of glycoprotein research in the future development of strategies on vector control to disrupt T. cruzi transmission.

A Risk Model Based on Ferroptosis-Related Genes OSMR, G0S2, IGFBP6, IGHG2, and FMOD Predicts Prognosis in Glioblastoma Multiforme.

Glioblastoma multiforme (GBM) is a common and highly aggressive brain tumor with a poor prognosis. However, the prognostic value of ferroptosis-related genes (FRGs) and their classification remains insufficiently studied. This study aims to explore the significance of ferroptosis classification and its risk model in GBM using multi-omics approaches and to evaluate its potential in prognostic assessment. Ferroptosis-related genes (FRGs) were retrieved from databases such as FerrDB. The TCGA-GBM and CGGA-GBM datasets were used as training and testing cohorts, respectively. Univariate Cox regression and LASSO regression analyses were performed to establish a risk model comprising five genes (OSMR, G0S2, IGFBP6, IGHG2, FMOD). A Meta-analysis of integrated TCGA and GTEx data was conducted to examine the differential expression of these genes between GBM and normal tissues. Key gene protein expression differences were analyzed using CPTAC and HPA databases. Single-cell RNA sequencing (scRNA-seq) analysis was employed to explore the cell type-specific distribution of these genes. The five-gene risk model demonstrated significant prognostic value in GBM. Meta-analysis revealed distinct expression patterns of the identified genes between GBM and normal tissues. Protein expression analysis confirmed these differences. scRNA-seq analysis highlighted the diverse distribution of these genes across different cell types, offering insights into their biological roles. The ferroptosis-based risk model provides valuable prognostic insights into GBM and highlights potential therapeutic targets, emphasizing the biological significance of ferroptosis-related genes in tumor progression.

Iristectorin A Ameliorates Cisplatin-Induced Hepatorenal Injury in Mice Through Modulation of the Nrf2/HO-1 Signaling Pathway.

Cisplatin (CIS) is a chemotherapeutic agent frequently used in cancer treatment. However, depending on the dosage and duration of use, CIS can lead to hepatotoxicity and nephrotoxicity. Iristectorin A (IRIS), a natural flavonoid, has been found to exhibit antioxidant and protective effects. In this paper, we scrutinized the effects and molecular mechanisms of the IRIS on CIS-induced liver and kidney damage in mice. IRIS administration alleviated CIS-induced elevations in AST, ALT, ALP, BUN, and creatinine levels by approximately 12%, 15%, 11%, 21%, and 15%, respectively. It also inhibited liver and kidney MDA levels by approximately 29% and 28%, while enhancing liver and kidney GSH, SOD, and CAT levels by 47%-60%, 85%-70%, and 90%-55%, respectively. IRIS enhanced liver and kidney mRNA expression levels of Nrf2 (by approximately 1.6- and 1.5-fold, respectively), HO-1 (by 1.5- and 1.5-fold, respectively), and Bcl-2 (by 1.5- and 1.4-fold, respectively). In addition, IRIS suppressed the mRNA expression levels of NF-κB (by 0.7- and 0.7-fold), TNF-α (by 0.7- and 0.7-fold), Bax (by 0.8- and 0.7-fold), and Cas-3 (by 0.9- and 0.7-fold). Protein expression analysis revealed that IRIS increased Nrf2 (by 1.5- to 1.2-fold) and Bcl-2 levels (by 1.3- to 1.7-fold), and reduced Bax (by 0.7- to 0.8-fold) and Cas-3 (by 0.8- and 0.8-fold) levels altered by CIS treatment. Moreover, IRIS administration prevented histopathological changes in the liver and kidney caused by CIS. Ultimately, IRIS was found to substantially mitigate CIS-induced hepatorenal injury by targeting oxidative stress, inflammation, and apoptosis through regulation of the Nrf2/HO-1 signaling pathway. Therefore, IRIS holds potential as a therapeutic adjuvant in the use of CIS.

KLF4 activates LATS2 to promote cisplatin sensitivity in ovarian cancer through DNA damage.

We aimed to investigate the role of large tumor suppressor kinase 2 (LATS2) in cisplatin (DDP) sensitivity in ovarian cancer. Bioinformatic analysis explored LATS2 expression, pathways, and regulators. Quantitative reverse transcription-PCR measured LATS2 and KLF4 mRNA levels. Dual-luciferase and chromatin immunoprecipitation assays confirmed their binding relationship. Cell viability, half maximal inhibitory concentration (IC 50 ) values, cell cycle, and DNA damage were assessed using CCK-8, flow cytometry, and comet assays. Western blot analyzed protein expression. The effect of LATS2 on the sensitivity of ovarian cancer to DDP was verified in vivo . LATS2 and KLF4 were downregulated in ovarian cancer, with LATS2 enriched in cell cycle, DNA replication, and mismatch repair pathways. KLF4, an upstream regulator of LATS2, bound to its promoter. Overexpressing LATS2 increased G1-phase cells, reduced cell viability and IC 50 values, and induced DNA damage. Silencing KLF4 alone showed the opposite effect on LATS2 overexpression. Knocking out LATS2 reversed the effects of KLF4 overexpression on cell viability, cell cycle, IC 50 values, and DNA damage in ovarian cancer cells. Inhibiting LATS2 inactivated the Hippo-YAP signaling pathway. In vivo experiments showed that overexpression of LATS2 enhanced the sensitivity of ovarian cancer to DDP. KLF4 activates LATS2 via DNA damage to enhance DDP sensitivity in ovarian cancer, providing a potential target for improving treatment outcomes.

Study and analysis of the use of bevacizumab in the treatment of breast cancer

This study aimed to explore the application of bevacizumab in breast cancer treatment. We reviewed the research progress and current status of bevacizumab, analyzed its mechanism of action, and introduced protein expression analysis to further investigate its therapeutic effect. Bevacizumab is an angiogenesis inhibitor that prevents tumor growth by binding to and blockingvascular endothelial growth factor (VEGF). Data were obtained derived from the United States Society of Clinical Oncology (ASCO), which employed reversed-phase protein array technology (RPPA) to analyze the expression levels of related proteins. This included data on 210 proteins from 55 CTx samples and 54 Bev + CTx samples. We conducted a follow-up analysis based on the existing studies of nine protein signatures with corresponding beta coefficients identified by Lasso regression. However, only the first eight proteins were analyzed due to insufficient data. Additionally, we analyzed four of the 210 proteins with high expression levels. Statistical analysis revealed differences in protein the expression levels between the CTx and Bev + CTx groups, though these differences were not statistically significant. Clustering results displayed protein expression data through heat maps, revealing a clustering relationship among several proteins. This study provides a reference for the application of bevacizumab in breast cancer treatment.

Comparative analysis of protein expression profiles with genotypes in the diagnosis of Inborn Errors of Immunity

Background Inborn Errors of Immunity (IEIs) are genetic diseases resulting from harmful genetic variations that hinder the proper functioning of the immune system. The broad range of IEIs involves multiple systems, presenting characteristics similar to allergies, autoimmune or inflammatory diseases, and malignancies. Given this complexity, there is an urgent need for a precise multi-parametric molecular diagnostic approach. Objective In this work, we demonstrated the effectiveness of accurate diagnosis by flow cytometry in patients with IEI by comparing genotype analysis with the expression levels of particular proteins and signaling activities. Methods We examined the expression levels or signaling activities of 28 cell surface and intracellular proteins using flow cytometry in a cohort of 352 patients and 189 healthy controls, in conjunction with genotype analysis for comparison. Results: We identified alterations in protein expression in 60 individuals, among them, 55 exhibited the presence of an underlying pathogenic mutation. Complete loss of protein expression was observed in seven patients, constituting 2% of the total, while reduced protein expression was noted in 35 patients (9%). Notably, despite mutations in the relevant genes, protein expression levels were normal in five patients (2%), in all investigated patients. 37% of patients had elevated signaling activity, and 17% were suggestive of a particular IEI diagnosis following protein expression analysis. Conclusion The correspondence between flow cytometry-based protein analyses and genotype facilitates a prompt diagnosis, providing patients with swift access to therapeutic options.

Isolation method of brain microvessels from small frozen human brain tissue for blood-brain barrier protein expression analysis

BackgroundProtein expression analysis of isolated brain microvessels provides valuable insights into the function of the blood-brain barrier (BBB). However, isolation of brain microvessels from human brain tissue, particularly in small quantities, poses significant challenges. This study presents a method for isolating brain microvessels from a small amount of frozen human brain tissue, adapting techniques from an established mouse brain capillary isolation method.MethodsBrain microvessel fractions were obtained from approximately 0.3 g of frozen human brain tissue (frontal cortex) using a bead homogenizer for homogenization, followed by purification with a combination of cell strainers and glass beads. Protein expression in the isolated human microvessel fractions and whole-brain lysates was analyzed by western blot and proteomic analysis.ResultsMicroscopic imaging confirmed the successful isolation of brain microvessels from frozen human brain tissue. Protein quantification assays demonstrated that the microvessel fraction yielded sufficient protein for detailed expression analysis. Western blot analysis revealed an enrichment of BBB-selective proteins including multidrug resistance 1 (MDR1)/ATP-binding cassette sub-family B member 1 (ABCB1), glucose transporter protein type 1 (GLUT1)/solute carrier family 2 member 1 (SLC2A1), and claudin 5 (CLDN5), in the brain microvessel fraction compared to whole-brain lysates. Multiple reaction monitoring quantification of six BBB-selective proteins—MDR1, breast cancer resistance protein (BCRP)/ATP binding cassette subfamily G member 2 (ABCG2), GLUT1, monocarboxylate transporter 1 (MCT1)/solute carrier family 16 member 1 (SLC16A1), transferrin receptor, and CLDN5—revealed expression levels consistent with those observed in larger human brain samples. Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH-MS)-based quantitative proteomics further demonstrated significant enrichment of human microvascular endothelial cells in the isolated fraction, corroborating the findings from mouse models.ConclusionsWe successfully developed a method for isolation of brain microvessels from a small amount of frozen human brain tissue, facilitating detailed study of BBB proteome in aging or pathological conditions. This technique provides valuable insights into BBB dysfunction in central nervous system disorders and holds potential for improving brain-targeted drug delivery strategies.

Tyrosine phosphatase SHP2 promoted the progression of CRC via modulating the PI3K/BRD4/TFEB signaling induced ferroptosis

ObjectiveTo elucidate the mechanism by which tyrosine phosphatase SHP2 protects CRC through modulation of TFEB-mediated ferritinophagy, thereby suppressing ROS and ferroptosis.MethodsSW480 and SW620 cells, in the logarithmic growth phase, were treated with or without the SHP2 inhibitor PHPS1, the activator Trichomide A, EGF, or MMP inhibitors, and randomly assigned to four groups. Additionally, SW480 cells in the logarithmic phase underwent treatments with EGF, the ferroptosis inducer erastin, Trichomide A, or the curcumin analog C1, forming seven groups. Cell migration assessment in these groups employed scratch and Transwell assays. Protein expression analysis of total SHP2, total PI3K, p-SHP2, p-PI3K, p-TFEB, TFEB, SQSTM1, LC3, LAMP2, NCOA4, FTH1, GPX4, NOX4, and ACSL4 in the seven SW480 groups was conducted through Western blot and immunofluorescence. Apoptosis analysis was performed on these seven groups, while gene co-expression analysis utilized bioinformatics. SW480 and CCD-841CoN cells were categorized into four groups, undergoing treatment with saline, EGFR-OE lentivirus, SHP2-KD lentivirus, or SHP2-OE lentivirus. Western blot analysis in SW480 cells detected EGFR, total SHP2, p-SHP2, GPX4, and ACSL4 proteins, and tumor volume observations were conducted in a nude mouse xenograft model. Western blot also evaluated total SHP2, p-SHP2, GPX4, and ACSL4 protein expression in CCD-841CoN cells.ResultsBioinformatics analysis revealed correlations between EGFR and SHP2, SHP2 and PIK3CA, SHP2 and MAPK1, BRK4 and HIF1A, HIF1A and NCOA4, as well as TFEB and FTH1. Scratch and Transwell assays showed that SHP2 diminishes the migratory capacity of SW480 and SW620 cells. Western blot and immunofluorescence demonstrated that EGFR activation of SHP2 markedly elevated p-TFEB levels while reducing TFEB protein expression. EGF stimulation enhanced the expression of FTH1, GPX4, NOX4, and ACSL4. Combined stimulation with EGF and SHP2 further amplified the expression of p-SHP2, p-TFEB, and NCOA4 while reducing TFEB, SQSTM1, LC3, and LAMP2. Erastin augmented FTH1, GPX4, NOX4, and ACSL4 expression while decreasing p-SHP2, p-TFEB, TFEB, SQSTM1, LC3, LAMP2, and NCOA4. TFEB activation suppressed p-SHP2, p-TFEB, NCOA4, FTH1, and GPX4 expression, while promoting TFEB, SQSTM1, LC3, LAMP2, NOX4, and ACSL4 expression. Apoptosis assays indicated that SHP2 activation decelerated apoptosis in SW480 cells, whereas erastin under EGF stimulation accelerated apoptosis, as did TFEB activation. Western blot results in SW480 cells displayed that overexpression of EGFR or SHP2 significantly increased total SHP2, p-SHP2, and GPX4 expression while decreasing ACSL4 levels. SHP2 knockdown decreased total SHP2, p-SHP2, and GPX4 expression, with an increase in ACSL4 expression. In CCD-841CoN cells, overexpression of EGFR or SHP2 resulted in a decrease in p-SHP2 and an increase in total SHP2, more pronounced with SHP2 overexpression, while GPX4 and ACSL4 levels remained stable. SHP2 knockdown led to reduced EGFR, total SHP2, p-SHP2, and GPX4 expression, without a significant impact on ACSL4 levels. The nude mouse xenograft model demonstrated that EGFR overexpression significantly increased tumor size, whereas SHP2 overexpression markedly decreased tumor volume. SHP2 knockdown resulted in significantly larger tumors.ConclusionSHP2 advances CRC progression by modulating TFEB-mediated ferritinophagy, suppressing ROS and ferroptosis. Targeting SHP2 presents a promising therapeutic strategy for CRC.

Plasma-Activated Medium Inhibited the Proliferation and Migration of Non-Small Cell Lung Cancer A549 Cells in 3D Culture.

Lung cancer is the most common type of malignant tumor worldwide. Plasma-activated medium (PAM) is an innovative cancer treatment method that has received considerable scientific attention. The objective of this study is to evaluate the effects of PAM on the anti-tumor characteristics of non-small cell lung cancer (NSCLC) cells in two-dimensional (2D) and three-dimensional (3D) cultures. The effects of PAM treatment on the proliferative and migratory capabilities of A549 cells in 2D and 3D cultures were assessed using MTT, migration, invasion assays, and cell cycle, respectively. The study also investigated the impact of PAM treatment on the changes in the content of intracellular and extracellular reactive species and analyzed protein expression using the Western Blot method. PAM treatment inhibited the viability, migration, and invasion abilities of A549 cells in both 2D and 3D cultures, suppressed the epithelial-mesenchymal transition (EMT) process, and downregulated the expression of the RAS/ERK signaling pathway, which effectively inhibited tumor spheroid formation. Additionally, the effect of PAM on A549 cells was mediated through ROS-induced oxidative reactions, and PAM treatment exhibited greater cytotoxicity in 2D culture compared to 3D culture. As compared to 2D, the 3D cell culture model provides a viable in vitro cell model for studying the mechanisms of PAM treatment in lung cancer. PAM represents an effective new treatment for NSCLC.

Stattic suppresses p‑STAT3 and induces cell death in T‑cell acute lymphoblastic leukemia.

The present study investigated the therapeutic potential of Stattic, a selective inhibitor of STAT3, in treating T‑cell acute lymphoblastic leukemia (T‑ALL). The effects of Stattic on cell viability, STAT3 phosphorylation, apoptosis and autophagy in T‑ALL cell lines, and on tumor growth in a xenograft mouse model of T‑ALL, were assessed. Methods, including the Cell Counting Kit‑8 assay for cell viability, propidium iodide/Annexin V staining for apoptosis detection, western blotting for protein expression analysis, and a xenograft mouse model for evaluating in vivo tumor growth, were employed. The results showed that Stattic effectively reduced cell viability in a dose‑dependent manner, with significant reductions observed at concentrations of 1.25 µM and above in CCRF‑CEM cells (IC50=3.188 µM) and at 2.5 µM and above in Jurkat cells (IC50=4.89 µM) after 24 h of treatment. Concurrently, Stattic significantly suppressed the expression of phosphorylated STAT3, indicating its mechanism of action as a STAT3 pathway inhibitor. Furthermore, Stattic treatment induced both apoptosis and autophagy in CCRF‑CEM and Jurkat cells, as evidenced by the respective upregulation of cleaved caspase‑3 and LC3B. In a xenograft mouse model of T‑ALL, Stattic markedly inhibited tumor growth, with the greatest effect occurring at the highest dose of 30 mg/kg. These results suggested that Stattic holds promise as a therapeutic agent in T‑ALL by modulating key pathways involved in cell survival and proliferation. In conclusion, Stattic exhibited a significant therapeutic potential for T‑ALL via a dose‑dependent reduction of cell viability, inhibiting STAT3 phosphorylation, and promoting both apoptotic and autophagic cell death; however, further studies are required before clinical application.

BIRC5 as a prognostic and diagnostic biomarker in pan-cancer: an integrated analysis of expression, immune subtypes, and functional networks.

BIRC5 (Survivin) is a crucial anti-apoptotic protein overexpressed in various cancers, promoting tumor growth and treatment resistance. This study investigates its expression across 33 cancer types and explores its diagnostic, prognostic, and immune-related significance. We analyzed RNA-seq data from TCGA and protein expression data from the Human Protein Atlas. Expression levels were compared between tumor and normal tissues. Correlations with molecular and immune subtypes were explored using TISIDB. Prognostic significance was evaluated through survival analysis, Cox regression, and ROC curve analysis. The PPI network was constructed using STRING. BIRC5 was significantly overexpressed in tumor tissues across 33 cancer types, with higher expression levels observed in tumors compared to normal tissues. The protein expression analysis revealed a similar trend. BIRC5 expression was significantly correlated with various molecular and immune subtypes in multiple cancer types. Survival analysis indicated that high BIRC5 expression was associated with poor prognosis across multiple cancers, including lung adenocarcinoma (LUAD) and kidney renal clear cell carcinoma (KIRC). ROC analysis showed that BIRC5 exhibited strong diagnostic potential, with high AUC values (>0.9) in several cancers. The PPI network analysis identified key interacting proteins involved in the cell cycle and tumor progression, further supporting BIRC5's role in cancer biology. Functional experiments in lung adenocarcinoma (LUAD) revealed that BIRC5 upregulation enhances cell proliferation, migration, and invasion, while its knockdown suppresses these activities. BIRC5 is a promising diagnostic and prognostic biomarker in multiple cancers. Its association with immune subtypes suggests a potential role in the tumor immune microenvironment. These findings support BIRC5 as a therapeutic target for cancer treatment.

Identification and Expression Profiling of LEA Gene Family in Olive (Olea europaea L.)

The olive tree (Olea europaea L.) is a vital crop in Mediterranean agriculture, frequently exposed to harsh drought conditions. Among the molecular mechanisms that confer drought tolerance, Late Embryogenesis Abundant (LEA) proteins play a central role. These hydrophilic proteins function in cellular protection during water deficit, preventing protein denaturation, stabilizing membranes, and scavenging reactive oxygen species. In this study, we performed a comprehensive phylogenetic and gene expression analysis of LEA proteins in olive. We identified LEA genes expressed across different tissues and conducted a differential expression analysis to assess their response to drought stress. A phylogenetic tree was constructed to classify LEA family members, and expression data was mapped onto the tree to link evolutionary conservation with functional responses. LEA proteins were classified into distinct subgroups (LEA_1-5, Dehydrin, SMP) to highlight their functional diversity. Additionally, a tissue-specific expression heatmap was generated to illustrate the spatial dynamics of LEA gene activity. Our results provide valuable insights into the molecular mechanisms of drought tolerance in olive and offer potential targets for genetic improvement to enhance resilience in olive cultivation.

Interleukin-6 promotes the epithelial mesenchymal transition in canine tonsillar squamous cell carcinoma cells

Canine oral squamous cell carcinoma (CoSCC) is often associated with suppurative inflammation. Metastasis of malignant tumors is one of the signs of major interest in oncology because it speaks of disease progression, where the involvement of interleukin-6 (IL-6) in cancer progression is so far unknown. Therefore, the aim of this study was the determination of the role of IL-6 in metastasis in CoSCC cells model through expression analysis of mRNA and protein using real-time PCR and western blotting and assessment of cell migration and invasion. The messenger RNA (mRNA) expression level of IL-6 was elevated in CoSCC tissues, and the IL-6 receptor protein was expressed in CoSCC cell lines. Furthermore, IL-6 levels were associated and showed negative correlation with survival time (rs = −0.92857) in dogs with tonsillar SCC. Recombinant canine IL-6 (rcIL-6) treatment promoted migration and invasion, in addition to increasing the viable cell number of the tonsillar SCC cell line (TSCCLN#6). Consistently, the protein expression of phosphorylated ERK1/2 and STAT3 and Fascin1 (FSCN1) was upregulated by treatment with rcIL-6 in a dose-dependent manner. Treatment with ERK or STAT3 inhibitors abolished the effects of rcIL-6, and the ERK inhibitor successfully downregulated the expression of FSCN1. In conclusion, IL-6 may be involved in tonsillar CoSCC invasion and metastasis through the activation of the mitogen-activated protein kinase and Janus tyrosine kinase/signal transducer and activator of transcription signaling.

Searching for Protein Off-Targets of Prostate-Specific Membrane Antigen-Targeting Radioligands in the Salivary Glands.

Background: Prostate specific membrane antigen (PSMA)-targeted radioligand therapies represent a highly effective treatment for metastatic prostate cancer. However, high and sustain uptake of PSMA-ligands in the salivary glands led to dose limiting dry mouth (xerostomia), especially with α-emitters. The expression of PSMA and histologic analysis couldn't directly explain the toxicity, suggesting a potential off-target mediator for uptake. In this study, we searched for possible off-target non-PSMA protein(s) in the salivary glands. Methods: A machine-learning based quantitative structure activity relationship (QSAR) model was built for seeking the possible off-target(s). The resulting target candidates from the model prediction were subjected to further analysis for salivary protein expression and structural homology at key regions required for PSMA-ligand binding. Furthermore, cellular binding assays were performed utilizing multiple cell lines with high expression of the candidate proteins and low expression of PSMA. Finally, PSMA knockout (PSMA-/-) mice were scanned by small animal PET/MR using [68Ga]Ga-PSMA-11 for in-vivo validation. Results: The screening of the trained QSAR model did not yield a solid off-target protein, which was corroborated in part by cellular binding assays. Imaging using PSMA-/- mice further demonstrated markedly reduced PSMA-radioligand uptake in the salivary glands. Conclusion: Uptake of the PSMA-targeted radioligands in the salivary glands remains primarily PSMA-mediated. Further investigations are needed to illustrate a seemingly different process of uptake and retention in the salivary glands than that in prostate cancer.

Resveratrol Can Differentiate Human Melanoma Stem-like Cells from Spheroids Treated With All-trans Retinoic Acid.

Stem-like cancer cells are believed to be the leading cause of therapy resistance in malignant melanoma (MM). All-trans retinoic acid (ATRA) differentiation therapy is considered a promising approach to eradicate stem-like cancer cells, but some melanoma cells are resistant to ATRA. This study aimed to examine whether resveratrol (RS), a natural polyphenol compound, could improve the response of MM stem-like cells to ATRA and explore the possible underlying mechanisms. MM stem-like cells were established from a spheroid model of A375 human MM cell line. The response to RES alone and in combination with ATRA, was examined through analysis of cancer stemness, cell viability, and protein expression. The stem-like cells showed resistance to the anticancer drug docetaxel; however, the combination of RES and ATRA augmented the effects of docetaxel. Accordingly, these combinatorial effects were associated with significant inhibition of the expression levels of stemness markers CD133, OCT4, CD271, and ABCG2. The tested combinations also led to a significant increase in melanocyte differentiation marker SOX9, while efficiently suppressing the dedifferentiation marker SOX10. Notably, RES alone effectively up-regulated retinoic acid receptor beta (RARβ) expression and down-regulated crucial mediators like DNMT1, polycomb-group proteins EZH2, and BMI-1, which mechanistically explain how RES enhanced the differentiation-inducing effects of ATRA. The resistance of MM stem-like cells to ATRA can be attenuated by RES and combined applications of ATRA and RES provide a promising strategy for MM treatment.

Comparative in situ characterization of erythroid cells found throughout the developing tongue tissue, circulation, and liver of fetal mice.

Erythroid cells contribute to embryonic organ development and adult tissue repair supplying oxygen to tissues. During mouse development, the primitive erythroid cells produced in the extraembryonic blood islands of the yolk sac begin to circulate as immature and nucleated erythroblasts with the onset of cardiac contractions around embryonic day 9.5 (E9.5). On the other hand, the definitive erythroid progenitors derived from the yolk sac and arterial vessels colonize the fetal liver, where they mature into small, enucleated definitive erythrocytes that are released into circulation at E11.5. In many cases, however, erythroblasts are also generated in situ during the development of tissue vasculature. We hypothesized that the properties of erythroid cells generated by peripheral tissue hematopoiesis may differ from those of contemporaneously circulating erythroid cells because hematopoiesis is spatiotemporally distinct from typical primitive and definitive hematopoiesis. Comparative in situ protein expression analyses of erythroid cells in developing tongue, circulation, and liver of fetal mice were performed at E12.5 and E14.5, using immunofluorescence staining of several marker proteins for erythroid and endothelial cells. At E12.5, irregularly elongated immature vascular endothelial cells, many of which were in contact with nucleated erythroblasts, were distributed in the developing tongue. In the three fetal locations examined (tongue, circulation, and liver), most erythroid cells at E12.5 expressed CD31, which is involved in cell-cell interactions; however, the expression was downregulated by E14.5. Interestingly, at E12.5, several erythroblasts strongly expressing CD31 were found in the tongue, but less abundant in the circulation and fetal liver. Nearly all erythroblasts in E12.5 fetuses expressed primitive erythroid cell-specific βH1-globin; however, those strongly expressing the embryonic globin were scattered, particularly in the tongue, fewer in the circulation and fetal liver. On the contrary, erythroid cells expressing adult β1-globin were scarcely found in the tongue, which is in stark contrast to those in the circulation and fetal liver, where nearly all erythroid cells weakly expressed β1-globin at the embryonic stage. At E14.5, the number of βH1-globin-expressing cells drastically decreased; however, it was still significantly higher in the tongue than in the circulation and fetal liver. In all three locations, β1-globin accumulation and the enucleation of erythroid cells progressed uniformly and rapidly from E12.5 to E14.5. The results indicate that primitive erythroblasts in the tongue highly express CD31, mature more slowly, and are replaced by definitive erythroid cells more slowly than those in the circulation and fetal liver. Primitive erythroblasts produced together with immature vascular endothelial cells during vasculogenesis in the developing tongue may perform functions other than oxygen supply, such as regulating vascular remodeling.