Exosomal transfer of miR-181b-5p confers senescence-mediated doxorubicin resistance via modulating BCLAF1 in breast cancer.

BACKGROUND: Doxorubicin resistance represents a major clinical challenge for treating patients with advanced breast cancer (BC). Exosomes, exchanging genetic cargo between heterogeneous populations of tumour cells, have been proposed to mediate drug resistance and cancer progression in other cancer types. However, their specific role in mediating doxorubicin resistance in BC remains unclear. Here, we demonstrate the important role of exosomal miR-181b-5p (exo-miR-181b-5p) in mediating doxorubicin resistance. METHODS: Small-RNA sequencing and bioinformatic analyses were used to screen miRNAs mediating doxorubicin resistance in BC, which were further verified by RT-qPCR. SA-ß-gal staining assays allowed us to measure cellular senescence. Exosomes from patients' serum before and after neoadjuvant chemotherapy were isolated for exo-miR-181b-5p quantification. RESULTS: Doxorubicin-resistant BC cell lines exhibited upregulated exosomal miR-181b-5p. Addition of exo-miR-181b-5p actively fused with recipient cells and transferred a drug-resistant phenotype. Overexpression of miR-181b-5p downregulated p53/p21 levels and inhibited doxorubicin-induced G1 arrest and senescence by suppressing BCLAF1 expression in vitro. Further, in vivo experiments showed treatment of exo-miR-181b-5p inhibitors exhibited superior tumour control and reversed the doxorubicin-resistance phenotype, accompanied with increased tumoral BCLAF1. CONCLUSION: Our data suggests exo-miR-181b-5p as a prognostic biomarker and a therapeutic potential for exo-miR-181b-5p inhibitors in the treatment of doxorubicin-resistant BC patients.

Peripheral Blood Leukocyte Detection Based on an Improved Detection Transformer Algorithm.

The combination of a blood cell analyzer and artificial microscopy to detect white blood cells is used in hospitals. Blood cell analyzers not only have large throughput, but they also cannot detect cell morphology; although artificial microscopy has high accuracy, it is inefficient and prone to missed detections. In view of the above problems, a method based on Fourier ptychographic microscopy (FPM) and deep learning to detect peripheral blood leukocytes is proposed in this paper. Firstly, high-resolution and wide-field microscopic images of human peripheral blood cells are obtained using the FPM system, and the cell image data are enhanced with DCGANs (deep convolution generative adversarial networks) to construct datasets for performance evaluation. Then, an improved DETR (detection transformer) algorithm is proposed to improve the detection accuracy of small white blood cell targets; that is, the residual module Conv Block in the feature extraction part of the DETR network is improved to reduce the problem of information loss caused by downsampling. Finally, CIOU (complete intersection over union) is introduced as the bounding box loss function, which avoids the problem that GIOU (generalized intersection over union) is difficult to optimize when the two boxes are far away and the convergence speed is faster. The experimental results show that the mAP of the improved DETR algorithm in the detection of human peripheral white blood cells is 0.936. In addition, this algorithm is compared with other convolutional neural networks in terms of average accuracy, parameters, and number of inference frames per second, which verifies the feasibility of this method in microscopic medical image detection.

Interaction and Kinetics Study of the Co-Gasification of High-solid Anaerobic Digestate and Lignite.

This study aims at investigating the interaction and kinetics behavior of the co-gasification of digestate and lignite. The co-pyrolysis performances of digestate and lignite blended by dry process were better than that blended by wet process, while the wet-blending process could improve the performance in co-gasification stage because of the larger pore diameter and pore volume. When anaerobic digestion (AD) time was 40 days, the synergistic interaction between digestate and lignite were the most remarkable based on the results of thermogravimetric analysis (TG) and the experiments in the lab-scale downdraft fixed bed gasifier. Kinetics study showed that the increase of AD time and the addition of digestate in lignite decreased the activation energy of the co-gasification reaction.

Surface Wetting-Driven Separation of Surfactant-Stabilized Water-Oil Emulsions.

Four fluorocarbon polymers including polytetrafluoroethylene and polyvinylidene fluoride were coated on a stainless steel felt to separate emulsified water droplets from ultralow sulfur diesel (ULSD) fuels. The original fuel treated with clay to remove additives was additized again with four known surfactants including pentaerythrityoleate, (octadecadienoic acid) dipolymer, (octadecadienoic acid) tripolymer, and monoolein individually. The different surfactants adsorbed on the fuel-water interface reduce the interfacial intension with different intensities. The separation efficiency at various surfactant concentrations was used to evaluate the coalescence effect exerted by these coatings. It was found the separation was both surfactant- and coating-dependent. A fluoro-polyurethane coating (FC1) stood out to counteract the adverse effect of all the surfactants. Solid free energy was then measured using acid-base and Kaelble-Uy adhesion theories for all the coatings, but its correlation with coalescence was not found at all. Coating aging in surfactant-additized fuel on the coating's water wettability was also examined to better understand how historical wetting affects separation. A tumbled model for fluorocarbons was identified that well-explained the continuous decline of the water contact angle on the FC1 coating in fuel. Subject to the challenge of the foreign environment, the fluoroalkyl chains of the polymer tilt to expose the carbonyl groups underneath, resulting in favored coalescence separation in the presence of surfactants.

CHIR99021 promotes self-renewal of mouse embryonic stem cells by modulation of protein-encoding gene and long intergenic non-coding RNA expression.

Embryonic stem cells (ESCs) can proliferate indefinitely in vitro and differentiate into cells of all three germ layers. These unique properties make them exceptionally valuable for drug discovery and regenerative medicine. However, the practical application of ESCs is limited because it is difficult to derive and culture ESCs. It has been demonstrated that CHIR99021 (CHIR) promotes self-renewal and enhances the derivation efficiency of mouse (m)ESCs. However, the downstream targets of CHIR are not fully understood. In this study, we identified CHIR-regulated genes in mESCs using microarray analysis. Our microarray data demonstrated that CHIR not only influenced the Wnt/ß-catenin pathway by stabilizing ß-catenin, but also modulated several other pluripotency-related signaling pathways such as TGF-ß, Notch and MAPK signaling pathways. More detailed analysis demonstrated that CHIR inhibited Nodal signaling, while activating bone morphogenetic protein signaling in mESCs. In addition, we found that pluripotency-maintaining transcription factors were up-regulated by CHIR, while several developmental-related genes were down-regulated. Furthermore, we found that CHIR altered the expression of epigenetic regulatory genes and long intergenic non-coding RNAs. Quantitative real-time PCR results were consistent with microarray data, suggesting that CHIR alters the expression pattern of protein-encoding genes (especially transcription factors), epigenetic regulatory genes and non-coding RNAs to establish a relatively stable pluripotency-maintaining network.

Water-soluble triarylboron compound for ATP imaging in vivo using analyte-induced finite aggregation.

Adenosine 5'-triphosphate (ATP) is a multifunctional molecule that participates in many important biological processes. Currently, fluorescence indicators for ATP with high performance are in demand. Reported herein is a novel water-soluble triarylboron compound which displays an apparent ATP-dependent fluorescence enhancement when dispersed in water. It can selectively recognize ATP from other bioactive substances in vitro and in vivo. The ATP-induced finite aggregation endows the indicator with appreciable photostability and superior tolerance to environmental electrolytes. This indicator has been successfully applied to the ATP imaging in NIH/3T3 fibroblast cells. The difference in the ATP levels within the membrane and cytosol is clearly visible.

Research on automatic matching of online mathematics courses and design of teaching activities based on multiobjective optimization algorithm.

The teaching of the optimization algorithm is a new kind of swarm intelligence optimization technique, which is superior in optimizing many simple functions. Still, it is not evident in processing some complex problems (group and teaching classification). Achieving automatic matching and knowledge transfer in online courses is imperative in mathematics education. This study proposes a design scheme MTCBO-LR (multiobjective capability optimizer-logistic regression), based on multitask optimization, which enables precise knowledge transfer and data interaction among many educators. It incorporates the standard TLBO algorithm to optimize, provides a variety of learning tactics for students at different stages of mathematics instruction, and is capable of adaptively adjusting these strategies in response to actual teaching needs. Experimental results on various datasets reveal that the proposed method enhances searchability and group diversity in various optimization extremes and outperforms similar methods in resolving to multitask teaching problems.

Molecular docking studies, anti-Alzheimer's disease, antidiabetic, and anti-acute myeloid leukemia potentials of narcissoside.

In this research, we explored their capacity for Narcissoside antioxidant and anticholinergic, antidiabetic, and anti-acute myeloid leukaemia. Narcissoside's antioxidant activities were elucidated by the use of various bioanalytical assays. Narcissoside's radical scavenging activities were evaluated by DPPH• and ABTS•+ scavenging activities. On the other hand, IC50 values were calculated for DPPH•, and ABTS•+ scavenging, acetylcholinesterase, and α-glucosidase inhibition effects of narcissoside. IC50 values narcissoside, as 11.54 nM for AChE and 65.58 nM for α-glucosidase were calculated with % Activity-[Inhibitory] graphs. Then, ADME/T analysis of narcissoside molecule was performed to calculate the drug becoming parameters.

USP51/PD-L1/ITGB1-deployed juxtacrine interaction plays a cell-intrinsic role in promoting chemoresistant phenotypes in non-small cell lung cancer.

BACKGROUND: Programmed death ligand 1 (PD-L1) has been demonstrated to facilitate tumor progression and therapeutic resistance in an immune-independent manner. Nevertheless, the function and underlying signaling network(s) of cancer cell-intrinsic PD-L1 action remain largely unknown. Herein, we sought to better understand how ubiquitin-specific peptidase 51 (USP51)/PD-L1/integrin beta-1 (ITGB1) signaling performs a cell-intrinsic role in mediating chemotherapeutic resistance in non-small cell lung cancer (NSCLC). METHODS: Western blotting and flow cytometry were employed for PD-L1 detection in NSCLC cell lines. Coimmunoprecipitation and pulldown analyses, protein deubiquitination assay, tissue microarray, bioinformatic analysis and molecular biology methods were then used to determine the significance of PD-L1 in NSCLC chemoresistance and associated signaling pathways in several different cell lines, mouse models and patient tissue samples. Ubiquitin-7-amido-4-methylcoumarin (Ub-AMC)-based deubiquitinase activity, cellular thermal shift and surface plasmon resonance (SPR) analyses were performed to investigate the activity of USP51 inhibitors. RESULTS: We provided evidence that cancer cell-intrinsic PD-L1 conferred the development of chemoresistance by directly binding to its membrane-bound receptor ITGB1 in NSCLC. At the molecular level, PD-L1/ITGB1 interaction subsequently activated the nuclear factor-kappa B (NF-κB) axis to elicit poor response to chemotherapy. We further determined USP51 as a bona fide deubiquitinase that targeted the deubiquitination and stabilization of the PD-L1 protein in chemoresistant NSCLC cells. Clinically, we found a significant direct relationship between the USP51, PD-L1 and ITGB1 contents in NSCLC patients with chemoresistant potency. The elevated USP51, PD-L1 and ITGB1 levels were strongly associated with worse patient prognosis. Of note, we identified that a flavonoid compound dihydromyricetin (DHM) acted as a potential USP51 inhibitor and rendered NSCLC cells more sensitive to chemotherapy by targeting USP51-dependent PD-L1 ubiquitination and degradation in vitro and in vivo. CONCLUSIONS: Together, our results demonstrated that the USP51/PD-L1/ITGB1 network potentially contributes to the malignant progression and therapeutic resistance in NSCLC. This knowledge is beneficial to the future design of advanced cancer therapy.

Socioeconomic and clinical factors affecting the proportion of breast conserving surgery in Chinese women with breast cancer.

Background: This study investigated the socioeconomic and clinical factors affecting the proportion of breast conserving surgery (BCS) in China, to improve the proportion and success rate of BCS in Chinese breast cancer patients. Methods: Six hundred and forty breast cancer patients treated with BCS were compared with 700 selected breast cancer patients (controls) treated with modified radical mastectomy (MRM) in Tianjin Medical University Cancer Institute and Hospital from January 2005 to January 2018. Patients' socioeconomic and clinical factors were collected through telephone interviews or face-to-face interviews. A total of 5,660 BCS patients were enrolled to analyze independent factors affecting initial positive margins. Chi-squared test and multiple logistic regressions were used to examine factors associated with BCS. The locoregional recurrence-free survival (LRRFS), distant metastasis-free survival (DMFS), and overall survival (OS) were calculated using the Kaplan-Meier method and the survival distribution between BCS and MRM groups was compared by log-rank test. Results: Breast cancer patients who were younger, lived in urban areas, had medical insurance, and higher levels of education and Personal income were more likely to choose BCS. We also observed that patients of Han nationality were more likely to choose BCS. Univariate analysis showed that the frozen section analysis (FSA) positive margin was significantly correlated with tumor distance from the nipple, preoperative magnetic resonance imaging (MRI) examination, T stage, pathological subtype, and lymphovascular invasion (LVI). Multivariate analysis showed the distance from the nipple, T stage, pathological subtype, and LVI, and no preoperative MRI examination were independent predictors of positive resection margins. Multivariate analysis of the correlation between MRI findings and positive resection margins revealed that tumor size, non-mass enhancement (NME), and malignant enhancement surrounding the tumor were independent predictors of positive resection margins. Conclusions: In China, socioeconomic factors largely influence the choice of surgical procedures for breast cancer patients. A gradual reduction in the influence of socioeconomic factors on the proportion of BCS is recommended. Furthermore, preoperative MRI should be encouraged in patients preparing for BCS. Clinicopathological characteristics and MRI findings are significantly associated with a positive resection margin in breast cancer patients.

Biochanin A inhibits lung adenocarcinoma progression by targeting ZEB1.

Lung adenocarcinoma is the major subtype of lung cancer, accounting for approximately 40% of lung cancers. During clinical treatment, the emergence of chemotherapy resistance seriously affects the effectiveness of treatment. Thus, finding new chemotherapeutic sensitizers is considered to be one of the effective solutions. Biochanin A, as a naturally occurring isoflavone, has been demonstrated to exhibit anticancer effects in various tumors. However, the potential mechanisms of Biochanin A to inhibit tumor development have not been clarified. In the present study, we found that the combinational treatment of cisplatin and Biochanin A exhibited strong synergistic repression on lung adenocarcinoma growth and progression in vitro and in vivo. Considering that epithelial-mesenchymal transition (EMT) is recognized to be associated with both chemoresistance and metastasis, we examined the EMT-related markers and found that Biochanin A could specifically inhibit the expression of ZEB1. Importantly, Biochanin A chemosensitizes lung adenocarcinoma and inhibits cancer cell metastasis by suppressing ZEB1. At the molecular level, Biochanin A affects the stability of ZEB1 protein through the deubiquitination pathway and thereby influences the progression of lung adenocarcinoma. In conclusion, our finding elucidates the potential efficacy of Bichanin A as a chemosensitizer and provides new strategy for the chemotherapy of advanced lung adenocarcinoma.

Rapid Determination of ß-Glucan Content of Hulled and Naked Oats Using near Infrared Spectroscopy Combined with Chemometrics.

The quantification of ß-glucan in oats is of immense importance for plant breeders and food scientists to develop plant varieties and food products with a high quantity of ß-glucan. However, the chemical analysis of ß-glucan is time consuming, destructive, and laborious. In this study, near-infrared (NIR) spectroscopy in conjunction with Chemometrics was employed for rapid and non-destructive prediction of ß-glucan content in oats. The interval Partial Least Square (iPLS) along with correlation matrix plots were employed to analyze the NIR spectrum from 700-1300 nm, 1300-1900 nm, and 1900-2500 nm for the selection of important wavelengths for the prediction of ß-glucan. The NIR spectral data were pre-treated using Savitzky Golay smoothening and normalization before employing partial least square regression (PLSR) analysis. The PLSR models were established based on the selection of wavelengths from PLS loading plots that present a high correlation with ß-glucan content. It was observed that wavelength region 700-1300 nm is sufficient for the satisfactory prediction of ß-glucan of hulled and naked oats with R2c of 0.789 and 0.677, respectively, and RMSE < 0.229.

Deciphering the performance of polo-like kinase 1 in triple-negative breast cancer progression according to the centromere protein U-phosphorylation pathway.

In general, the lack of effective therapeutic targets has led to the poor prognosis of triple-negative breast cancer (TNBC). Polo-like kinase 1 (PLK1) has been studied extensively as an effective therapeutic objective for the progression of tumor. Although the fundamental strategy and function of PLK1 in TNBC are still unclear. Here, we demonstrated that PLK1 upregulation was significantly correlated with poor prognosis in breast cancer cases utilizing the TCGA database. Additionally, ectopic PLK1 expression promoted TNBC cell proliferation, VEGFA production, and endothelial cell tube formation, whereas PLK1 knockdown induced the opposite effects. Moreover, expression of PLK1 K82R, the kinase-dead mutant of PLK1, completely inhibited PLK1-mediated cell proliferation, VEGFA production, and tube formation. Gene Set Enrichment Analysis (GSEA) showed that PLK1 expression significantly correlated with mitosis and the VEGF signaling pathway. We further observed that PLK1 phosphorylated centromere protein U (CENPU) at residue T78, thereby regulating the signaling pathway of COX-2/HIF-1α/VEGFA and the metaphase-anaphase transition of mitosis. The mechanism underlying the activity of PLK1 was also determined using a TNBC xenograft mouse model. Moreover, a PLK1 inhibitor effectively inhibited TNBC progression. Taken together, our results revealed that PLK1 plays an important role in TNBC progression via its kinase activity and phosphorylation of CENPU. Thus, PLK1 is an effective therapeutic objective for TNBC.

Chidamide Combined With Doxorubicin Induced p53-Driven Cell Cycle Arrest and Cell Apoptosis Reverse Multidrug Resistance of Breast Cancer.

The multidrug-resistant (MDR) phenotype is usually accompanied by an abnormal expression of histone deacetylase (HDAC). Given that HDAC is vital in chromatin remodeling and epigenetics, inhibiting the role of HDAC has become an important approach for tumor treatment. However, the effect of HDAC inhibitors on MDR breast cancer has not been elucidated. This study aim to demonstrate the potential of chidamide (CHI) combined with the chemotherapy drug doxorubicin (DOX) to overcome chemotherapeutic resistance of breast cancer in vitro and in vivo, laying the experimental foundation for the next clinical application. The results showed that, CHI combined with DOX showed significant cytotoxicity to MDR breast cancer cells in vitro and in vivo compared with the CHI monotherapy. The cell cycle distribution results showed that CHI caused G0/G1 cell cycle arrest and inhibited cell growth regardless of the addition of DOX. At the same time, annexin V staining and TUNEL staining results showed that CHI enhanced the number of cell apoptosis in drug-resistant cells. The western blot analysis found that p53 was activated in the CHI-treated group and combined treatment group, and then the activated p53 up-regulated p21, apoptosis regulator recombinant protein (Puma), and pro-apoptotic protein Bax, down-regulated the apoptotic proteins Bcl-xL and Bcl-2, and activated the caspase cascade to induce apoptosis.

Over-Expression of Centromere Protein U Participates in the Malignant Neoplastic Progression of Breast Cancer.

The expression of Centromere Protein U (CENP-U) is closely related to tumor malignancy. Till now, the role of CENP-U in the malignant progression of breast cancer remains unclear. In this study, we found that CENP-U protein was highly expressed in the primary invasive breast cancer tissues compared to the paired adjacent histologically normal tissues and ductal carcinoma in situ (DCIS) tissues. After CENP-U was knocked down, the proliferation and colony-forming abilities of breast cancer cells were significantly suppressed, whereas the portion of apoptotic cells was increased. Meanwhile, the PI3K/AKT/NF-κB pathway was significantly inhibited. In vivo studies showed that, the inhibition of CENP-U repressed the tumor growth in orthotopic breast cancer models. Therefore, our study demonstrated that the CENP-U might act as an oncogene and promote breast cancer progression via activation of the PI3K/AKT/NF-κB pathway, which suggests a promising direction for targeting therapy in breast cancer.

Chemotherapy-elicited exosomal miR-378a-3p and miR-378d promote breast cancer stemness and chemoresistance via the activation of EZH2/STAT3 signaling.

BACKGROUND: Not all breast cancer (BC) patients who receive neoadjuvant chemotherapy achieve a pathologic complete response (pCR), but the reasons for this are unknown. Previous studies have shown that exosomes produced in the tumor microenvironment in response to chemotherapy promote a chemotherapy-resistant phenotype in tumors. However, the role of BC chemotherapy-elicited exosomes in regulating chemoresistance is poorly understood. METHODS: Using commercial kits, serum exosomes were extracted from patients before neoadjuvant chemotherapy, after one cycle of chemotherapy and after four cycles of chemotherapy consisting of doxorubicin (DOX) and paclitaxel (PTX). Their miRNAs were sequenced, and the correlation between the sequencing results and chemotherapy effects was further verified by RT-qPCR using patient serum exosomes. Cell Counting Kit-8 (CCK-8) was used to detect chemosensitivity. Stemness was assessed by CD44+/CD24- population analysis and mammosphere formation assays. Chromatin immunoprecipitation (ChIP) experiments were performed to verify the binding of signal transducer and activator of transcription 3 (STAT3) to the promoter of miRNAs. RESULTS: Here, we provide clinical evidence that chemotherapy-elicited exosomal miR-378a-3p and miR-378d are closely related to the chemotherapy response and that exosomes produced by BC cells after stimulation with DOX or PTX deliver miR-378a-3p and miR-378d to neighboring cells to activate WNT and NOTCH stemness pathways and induce drug resistance by targeting Dickkopf 3 (DKK3) and NUMB. In addition, STAT3, which is enhanced by zeste homolog 2 (EZH2), bound to the promoter regions of miR-378a-3p and miR-378d, thereby increasing their expression in exosomes. More importantly, chemotherapeutic agents combined with the EZH2 inhibitor tazemetostat reversed chemotherapy-elicited exosome-induced drug resistance in a nude mouse tumor xenograft model. CONCLUSION: This study revealed a novel mechanism of acquired chemoresistance whereby chemotherapy activates the EZH2/STAT3 axis in BC cells, which then secrete chemotherapy-elicited exosomes enriched in miR-378a-3p and miR-378d. These exosomes are absorbed by chemotherapy-surviving BC cells, leading to activation of the WNT and NOTCH stem cell pathways via the targeting of DKK3 and NUMB and subsequently resulting in drug resistance. Therefore, blocking this adaptive mechanism during chemotherapy may reduce the development of chemotherapy resistance and maximize the therapeutic effect.

Normal red blood cell count reference values in Chinese presenile women given by geographical area.

BACKGROUND/PURPOSE: We aimed to standardize the normal reference value of red blood cell (RBC) counts in Chinese presenile women using an underlying scientific basis. METHODS: This research was conducted to study the relationship between the normal reference value of 31,405 RBC samples from presenile women in eight different geographical areas in China. RBC counts were determined using a microscopic counting method. RESULTS: There was a significant correlation between geographical factors and the normal reference RBC value in presenile women (F = 187.82, p = 0.000). Using stepwise regression analysis, one regression equation was obtained. CONCLUSION: If geographical values are obtained in a certain area, the normal RBC reference value in presenile women in this area can be obtained using the regression equation.

Amidoximated cellulose fiber membrane for uranium extraction from simulated seawater.

Uranium extraction from seawater is considered as an efficient strategy to meet the increasing demands of uranium. Amidoxime has been reported as one of the most efficient groups for uranium affinity. Herein, amidoximated cellulose fibers were synthesized by grafting polyacrylonitrile (PAN) onto cellulose fibers followed by amidoxime modification. The amidoximated cellulose fibers showed maximum adsorption capacity of 52.88 mg g-1 (pH = 5.0), and its static adsorption process was well fitted with Langmuir model and Pseudo-second-order kinetics. The adsorption mechanism was attributed to the chelating reaction between uranyl complexes and amidoximated cellulose fibers. The prepared fibers were further fabricated into nonwoven membrane for dynamic adsorption, and the breakthrough curves were well fitted to Dose-Response model. The amidoximated cellulose fiber membrane showed a good adsorption capacity of 1.22 mg g-1 at pH 8.0 after filtrating 10.0 L simulated seawater, demonstrating promising efficient engineering materials for uranium extraction from seawater.

Centromere protein U (CENPU) enhances angiogenesis in triple-negative breast cancer by inhibiting ubiquitin-proteasomal degradation of COX-2.

Triple-negative breast cancer (TNBC) is characterized by high vascularity, but anti-angiogenic therapies show poor efficacy. Centromere protein U (CENPU), a centromere component essential for mitosis, is associated with tumorigenesis in multiple cancers; however, little is known of its role in breast cancer. Here, we investigate its expression and function of promoting angiogenesis in TNBC. Immunohistochemical staining revealed high CENPU expression in TNBC tissue and high CENPU levels correlated significantly with poor distant metastasis-free and overall survival. Knockdown of CENPU in TNBC cells inhibited vascular endothelial growth factor A (VEGFA) production and significantly reduced tube formation and migration of human umbilical vein endothelial cells in vitro. In a mouse xenograft model, CENPU knockdown reduced TNBC tumor growth concomitant with a reduction in CD31 + microvessel density. Mechanistic studies revealed that CENPU promoted angiogenesis by inhibiting the ubiquitination and proteasomal degradation of cyclooxygenase-2 (COX-2), leading to increased activation of the COX-2-p-ERK-HIF-1α-VEGFA signaling pathway. Taken together, our results demonstrate a critical role for CENPU in COX-2-mediated signaling for angiogenesis, and identify a role of CENPU in regulating angiogenesis in TNBC.

Single-cell RNA-seq unveils critical regulators of human FOXP3+ regulatory T cell stability.

The heterogeneity and plasticity of T lymphocytes is critical for determining immune response outcomes. Functional regulatory T (Treg) cells are commonly characterized by stable FOXP3 expression and have reported to exhibit heterogeneous phenotypes under inflammatory conditions. However, the interplay between inflammation and Treg cell suppressive activity still remains elusive. Here, we utilized single-cell RNA sequencing to investigate how human Treg cells respond to the pro-inflammatory cytokine interleukin-6 (IL-6). We observed that Treg cells divided into two subpopulations after IL-6 stimulation. TIGIT- unstable Treg cells lost FOXP3 expression and gained an effector-like T cell phenotype, whereas TIGIT+ Treg cells retained robust suppressive function. Single cell transcriptome analysis revealed a spectrum of cellular states of IL-6-stimulated Treg cells and how cytochrome P450 family 1 subfamily A member 1 (CYP1A1) is a crucial regulator of Treg cell suppressive capability and stability. CYP1A1-deficient human Treg cells developed a Th17-like phenotype after IL-6 stimulation. Our findings implicate CYP1A1 as a previously unidentified regulator of Treg cells that may have target potential for clinical application for biotherapies.