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Risk factors for hepatocarcinogenesis include chronic inflammation due to viral infection, liver fibrosis, and aging. In this study, we separated carcinogenic and non-carcinogenic cases due to hepatitis C virus (HCV) infection, aiming to comprehensively analyze miRNA expression in liver tissues by age, and identify factors that contribute to carcinogenesis. Total RNA was extracted from 360 chronic hepatitis C (CH), 43 HCV infected hepatocellular carcinoma (HCC), and surrounding non-tumor (SNT) tissues. MicroRNA (miRNA) expression patterns were analyzed using microarray. Using machine learning, we extracted characteristic miRNA expression patterns for each disease and age. There were no age-dependent changes in miRNA expression in the disease-specific comparisons; however, miRNA expression differed among the age groups of 50, 60, and 70 years of age between CH and SNT. The expression of miRNA was different between SNT and HCC only in patients in their 70s. Of the 55 miRNAs with significant differences in expression between CH and SNT, 34 miRNAs showed significant differences in expression even in the degree of liver fibrosis. The observation that miRNAs involved in hepatocarcinogenesis differ at different ages suggests that the mechanisms of carcinogenesis differ by age group as well. We also found that many miRNAs whose expression did not affect liver fibrosis were involved in carcinogenesis. These findings are expected to define biomarkers for detection of HCC at early stage, and develop novel therapeutic targets for HCC.
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Carcinoma Hepatocelular , Hepatitis C Crónica , Neoplasias Hepáticas , MicroARNs , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/virología , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/virología , Persona de Mediana Edad , Masculino , Femenino , Anciano , Hepatitis C Crónica/genética , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/patología , Cirrosis Hepática/genética , Cirrosis Hepática/virología , Cirrosis Hepática/patología , Hígado/metabolismo , Hígado/patología , Hígado/virología , Perfilación de la Expresión Génica , Carcinogénesis/genética , Adulto , Regulación Neoplásica de la Expresión GénicaRESUMEN
Introduction: In the evolving healthcare landscape, we aim to integrate hyperspectral imaging into Hybrid Health Care Units to advance the diagnosis of medical diseases through the effective fusion of cutting-edge technology. The scarcity of medical hyperspectral data limits the use of hyperspectral imaging in disease classification. Methods: Our study innovatively integrates hyperspectral imaging to characterize tumor tissues across diverse body locations, employing the Sharpened Cosine Similarity framework for tumor classification and subsequent healthcare recommendation. The efficiency of the proposed model is evaluated using Cohen's kappa, overall accuracy, and f1-score metrics. Results: The proposed model demonstrates remarkable efficiency, with kappa of 91.76%, an overall accuracy of 95.60%, and an f1-score of 96%. These metrics indicate superior performance of our proposed model over existing state-of-the-art methods, even in limited training data. Conclusion: This study marks a milestone in hybrid healthcare informatics, improving personalized care and advancing disease classification and recommendations.
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Cysteine and serine-rich nuclear protein 1 (CSRNP1) has shown prognostic significance in various cancers, but its role in non-small cell lung cancer (NSCLC) remains elusive. We investigated CSRNP1 expression in NSCLC cases using bioinformatics tools from the GEO public repository and validated our findings through RT-qPCR in tumor and adjacent normal tissues. KEGG and GO enrichment analyses were employed to unveil the significant deregulation in signaling pathways. Additionally, clinical significance of CSRNP1 in NSCLC was determined through receiver operating curve (ROC) analysis, and its impact on survival was assessed using Kaplan-Meier analysis. To explore the functional impact of CSRNP1, we silenced its expression in NSCLC cells and assessed the effects on cell viability, migration, and invasion using MTT, Transwell, and wound-healing assays, respectively. Additionally, we investigated the influence of CSRNP1 silencing on the phosphorylation patterns of critical signaling proteins such as p53, p-Akt, and p-MDM2. Our results demonstrated significantly lower CSRNP1 expression in NSCLC tumor tissues (P < 0.01). ROC analysis indicated that NSCLC patients with high CSRNP1 expression exhibited extended overall survival and disease-free survival. Furthermore, CSRNP1 silencing promoted NSCLC cells viability, migration, and invasion (P < 0.05). Mechanistically, CSRNP1 silencing led to increased phosphorylation of AKT and MDM2, along with a concurrent reduction in p53 protein expression, suggesting its impact on NSCLC through deregulated cell cycle processes. In conclusion, our study underscores the significance of CSRNP1 in NSCLC pathogenesis, offering insights for targeted therapeutic interventions of NSCLC.
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Objective To investigate the expression and potential diagnostic value of six-transmembrane epithelial antigen of prostate 1(STEAP1)in bladder transitional cell carcinoma.Methods 52 patients with transitional cell carcinoma of the bladder who underwent surgical treatment at the 940th Hospital of Joint Lo-gistics Support Force of the Chinese People's Liberation Army from June 2021 to December 2022 were select-ed as the observation group.In addition,52 patients with benign tumors of the bladder who matched basic clin-ical data such as age,gender,and disease incidence were selected as the control group.The relative expression levels of STEAP1 and STEAP1 mRNA in bladder tumor tissues of patients in the two groups were deter-mined by enzyme-linked immunosorbent assay and real-time fluorescence quantitative PCR,and the relative expression levels of STEAP1 and STEAP1 mRNA in bladder tumor tissues of patients with different patho-logical parameters were compared.Spearman correlation analysis and binary Logistic regression analysis were used to screen the risk factors for the occurrence and clinical stage of bladder transitional cell carcinoma.Re-ceiver operating characteristic(ROC)curve and area under the curve(AUC)were used to evaluate the diagnos-tic and predictive value of each indicator for bladder transitional cell carcinoma.Results The relative expres-sion levels of STEAP1 and STEAP1 mRNA in bladder tumor tissues in observation group were significantly higher than those in control group,with statistical significance(P＜0.05).The relative expression levels of STEAP1 and STEAP1 mRNA in bladder tumor tissues of patients with middle and advanced bladder transi-tional cell carcinoma were significantly higher than those of patients with early bladder transitional cell carci-noma,with statistical significance(P＜0.05).Binary Logistic regression analysis showed that the relative ex-pression level of STEAP1 and STEAP1 mRNA in bladder tumor tissues of patients were independent risk fac-tors for the development of bladder transitional cell carcinoma and middle and advanced bladder transitional cell carcinoma(P＜0.05).ROC curve analysis showed that the AUC of STEAP1 and STEAP1 mRNA inde-pendently predicting the occurrence of bladder transitional cell carcinoma was 0.841(95％CI:0.760-0.922,P＜0.001)and 0.936(95％CI:0.893-0.980,P＜0.001),respectively,both of which had high predictive ef-ficacy.Conclusion The relative expression levels of STEAP1 and STEAP1 mRNA in bladder tumor tissues of patients are positively correlated with the occurrence of bladder transitional cell carcinoma and the middle and advanced bladder transitional cell carcinoma,suggesting that STEAP1 can be used as a potential marker for di-agnosis and prediction of the occurrence and development of bladder transitional cell carcinoma.
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BACKGROUND: Surgical resection followed by indicated adjuvant therapy offers potential curative treatment in colonic adenocarcinoma. Beyond the well-established seed and soil theory of colon cancer progression, the 'normal-appearing' tissues near the tumor are not genuinely normal and remain as remnants in patients following surgery. Our objective was to elucidate the alteration of gene expression and pathways across various distances of resection margins in right-sided colonic adenocarcinoma. METHODS: Twenty-seven fresh samples of primary cancer and 56 matched non-tumor tissues adjacent to the tumor (NAT) were collected from patients with resectable right-sided colon cancer. NAT were systematically obtained at varying distances (1, 5, and 10 cm) on both proximal and distal sides. Comprehensive gene expression analysis was performed using 770-gene PanCancer Progression Panel, delineating distinctive pathways and functional predictions for each region. RESULTS: Distinctive gene signatures and pathways exhibited by normal-appearing tissues were discovered at varying distances from cancer. Notably, SFRP2, PTGDS, COL1A1, IL1B, THBS2, PTGIS, COL1A2, NPR1, and BGN were upregulated, while ENPEP, MMP1, and NRCAM were downregulated significantly in 1-cm tissue compared to farther distances. Substantial alterations in the extracellular matrix (ECM) and prostaglandin/thromboxane synthesis were significantly evident at the 1-cm distance. Functional analysis indicated enhanced cell viability and survival, alongside reduced cellular death and apoptosis. CONCLUSIONS: Different distances exerted a significant impact on gene alteration within the normal-looking mucosa surrounding primary cancer, influenced by various mechanisms. These findings may highlight potential therapeutic targets related to the ECM and prostaglandin/thromboxane pathways for treatment strategies.
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Adenocarcinoma , Neoplasias del Colon , Humanos , Prostaglandinas , Márgenes de Escisión , Adenocarcinoma/genética , Adenocarcinoma/cirugía , Adenocarcinoma/patología , Neoplasias del Colon/genética , Neoplasias del Colon/cirugía , Matriz Extracelular/genética , Matriz Extracelular/patología , TromboxanosRESUMEN
Tumor selective near-infrared (NIR) fluorescent contrast agents has the potential to greatly enhance the efficiency and precision of tumor surgery by enabling real-time tumor margin identification for tumor resection guided by imaging. However, the development of these agents is still challenging. In this study, based on the acidic tumor microenvironment (TME), we designed and synthesized a novel pH-sensitive NIR fluorescent contrast agent OBD from ß-carboline. The fluorescence quantum yield of OBD exhibited a notable increase at pH 3.6, approximately 12-fold higher compared to its value at pH 7.4. After cellular uptake, OBD lighted up the cancer cells with high specificity and accumulated in the mitochondria. Spraying OBD emitted selective fluorescence in xenograft tumor tissues with tumor-to-normal tissue ratios (TNR) as high as 11.18, implying successful image-guided surgery. Furthermore, OBD was also shown to track metastasis in spray mode. After simple topical spray, OBD rapidly and precisely visualized the tumor margins of clinical colon and liver tissues with TNR over 4.2. Therefore, the small-molecule fluorescent contrast agent OBD has promising clinical applications in tumor identification during surgery.
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Medios de Contraste , Neoplasias , Humanos , Microambiente Tumoral , Neoplasias/diagnóstico por imagen , Colorantes Fluorescentes/química , Fluorescencia , Imagen Óptica/métodosRESUMEN
The well-known small-molecule biothiols have been used to maintain the normal metabolism of peroxy radicals, forming protein structures, resisting cell apoptosis, regulating metabolism, and protecting the homeostasis of cells in the organism. A large amount of research has found that abnormal levels of the above biothiols can cause some adverse diseases, such as changes in hair pigmentation, a slower growth rate, delayed response, excessive sleep and skin diseases. In order to further investigate the exact intracellular molecular mechanism of biothiols, it is imperative to explore effective strategies for real-time biothiol detection in living systems. In this work, a new near-infrared (NIR) emission fluorescence probe (probe 1) for sensitive and selective detection of biothiols was devised by combining dicyanoisophorone derivatives with the dinitrobenzenesulfonyl (DNBS) group. As expected, probe 1 could specifically detect biothiols (Cys, Hcy and GSH) through the dinitrobenzenesulfonyl group to form dye 2, which works as a signaling molecule for sensing biothiols in real samples. Surprisingly, probe 1 showed superior sensing characteristics and low-limit detection towards biothiols (36.0 nM for Cys, 39.0 nM for Hcy and 48.0 nM for GSH) with a large Stokes shift (134 nm). Additionally, the function of probe 1 as a platform for detecting biothiols was confirmed by confocal fluorescence imaging of biothiols in MCF-7 cells and zebrafish. More importantly, the capability of probe 1 in vivo has been further evaluated by imaging the overexpressed biothiols in tumor tissue. It is reasonable to believe that probe 1 can provide a valuable method to explore the relationship between biothiols and the genesis of tumor.
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Rationale: Challenges such as developing a universal tumor-specific probe for tumor margin identification in diverse tumors with an easy-operative and fast-imaging pattern still exist. Hence, in the present study, a rapidly "off-on" near-infrared (NIR) fluorescent probe NBD with pH-activatable fluorescence and a large Stokes shift was constructed for spray mediated near-instant and precise clinical tumor margins identification. Methods: NBD was designed and synthesized by introducing both diphenyl amino group and benzo[e]indolium to ß-carboline at C-6 and C-3 positions respectively. The optical properties of NBD was characterized by absorption spectra, fluorescence spectra. Subsequently, we investigated its pH-dependent mechanism by 1H NMR and density functional theory (DFT) calculation. NBD was further under deeper investigation into its imaging performance in nude mice models (subcutaneous, orthotopic, metastatic tumor), and clinical tissues from patients with three clinically representative tumors (liver cancer, colon cancer, and lung cancer). Results: It was found that NBD had NIR fluorescence (742 nm), a large Stokes shift (160 nm), and two-photon absorbance (1040 nm). Fluorescence quantum yield (ФF) increased by 5.5-fold when pH decreased from 7.4 to 4.0, to show pH-dependent property. Furthermore, NBD could not only selectively light up all four cancer cell lines, but also delineate xenograft tumor and orthotopic microtumor to guide surgical tumor resection, and track metastatic tissues. Particularly, after simple topical spray (three minutes later), NBD could rapidly and precisely distinguish the boundary ranges of three kinds of clinical cancer specimens including liver, colon, and lung cancers, with high tumor-to-normal tissue signal ratios (6.48~9.80). Conclusions: Therefore, the proposed fluorescent probe NBD may serve as a versatile NIR fluorogenic spray for the near-instant visualization of tumor margins and assisting surgeons in surgerical resection of clinical cancers.
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Neoplasias del Colon , Neoplasias Pulmonares , Animales , Ratones , Humanos , Colorantes Fluorescentes , Ratones Desnudos , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/cirugía , Concentración de Iones de HidrógenoRESUMEN
Background: We investigated the expression pattern of a human stem cell-specific, large intergenic noncoding RNA (lincRNA) regulator of reprogramming (lincRNA-ROR) and its spliced transcript variants in breast tumors. Breast cancer is the leading cause of cancer mortality in women; therefore, finding a reliable diagnostic tumor marker, based on the molecular profile of tumor cells, is warranted. Methods: qRT-PCR was used to investigate the expression alteration of a specific stem cell-related lincRNA and its spliced transcript variants in breast tumors which provided by the Iran National Tumor Bank (2014-2016). Suitability of lincRNA-ROR and expression alterations of its spliced transcript variants as breast tumor biomarkers were examined by ROC curve analysis. Results: Expression was significantly upregulated in lincRNA-ROR variants 1 (NR-048536) and 4 (AB844432) and downregulated in variant 3 (AB844431), with expression levels failing to distinguish between breast tumor types, grades, and malignancy stages. Whereas ROC curve analysis gave good scores to the expressions of variants 1 (AUC=0.7675, P=0.003) and 3 (AUC=0.9383, P=0.00173), suggesting their suitability as potential breast tumor biomarkers, it gave an AUC score of 0.6033 for lincRNA-ROR spliced variant 4 (P=0.4118), denoting its unsuitability as a breast cancer biomarker. Conclusion: Aberrant expressions of lincRNA-ROR spliced transcript variants could serve as reliable biomarkers with potential usefulness in breast cancer diagnosis. However, further research should elucidate the role and tissue expression of lincRNA-ROR spliced transcript variants in various cancers.
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Patient-derived organoids (PDO) are a new biomedical research model that can reconstruct phenotypic and genetic characteristics of the original tissue and are useful for research on pathogenesis and drug screening. To introduce the progression in this field, we review the key factors of constructing organoids derived from epithelial tissues and cancers, covering culture medium and matrix, morphological characteristics, genetic profiles, high-throughput drug screening, and application potential. We also discuss the co-culture system of cancer organoids with tumor microenvironment (TME) associated cells. The co-culture system is widely used in evaluating crosstalk of cancer cells with TME components, such as fibroblasts, endothelial cells, immune cells, and microorganisms. The article provides a prospective for standardized cultivation mode, automatic morphological evaluation, and drug sensitivity screening using high-throughput methods.
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Células Endoteliales , Neoplasias , Humanos , Evaluación Preclínica de Medicamentos , Estudios Prospectivos , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Neoplasias/patología , Organoides/patología , Microambiente TumoralRESUMEN
Herein, a novel fluorescent probe, GTP, was developed for monitoring the GGT (γ-glutamyl transpeptidase) level in living cells and biopsies. It consisted of the typical recognition group γ-Glu (γ-Glutamylcysteine) and the fluorophore (E)-4-(4-aminostyryl)-1-methylpyridin-1-ium iodide. With a ratio response between the signal intensity at 560 nm and 500 nm (RI560/I500), it could be important complement for the turn-on ones. With the linear range of 0-50 U/L, the limit of detection was calculated as 0.23 µM. The detection system showed the strongest response near pH 7.4, and exhibited steady fluorescence signals for at least 48 h. With high selectivity, good anti-interference and low cytotoxicity, GTP was suitable for physiological applications. By monitoring the GGT level with the ratio values in the green and blue channels, the probe GTP could distinguish cancer cells from normal cells. Furthermore, in the mouse tissues and humanization tissue samples, the probe GTP could also recognize the tumor tissues from the normal ones.
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Colorantes Fluorescentes , gamma-Glutamiltransferasa , Animales , Ratones , Colorantes Fluorescentes/toxicidad , Diagnóstico por Imagen , Biopsia , Guanosina TrifosfatoRESUMEN
Background: RNA integrity of tumor tissues from 12 common organs was measured, and tumor tissues from liver were found to have the best RNA integrity in our previous study. The effects of preanalytical variables in the phase of pre- and postacquisition on RNA integrity were further assessed in hepatocellular carcinoma (HCC) tissues in this study. Methods: RNA integrity number (RIN) was measured in tissues from 146 HCC patients. First, 42 fresh HCC tumor tissues were newly collected to assess the effect of various preanalytical variables in the phase of preacquisition on RNA integrity. Second, eight paired HCC tumor and normal tissues were newly collected and used in the gradient course study of ex vivo ischemia time and freeze-thaw cycles on RNA integrity. Finally, 96 stock-frozen tumor tissues with various years of frozen storage were used to assess the effect of cryopreservation time. Results: RNA integrity was found to be independent of patient age, sex, clinical stage, tumor location, HBV infection status, tumor diameter, and surgical approach, but affected by tumor grade. Tumor tissues with a greater tumor grade had lower RIN. With the prolongation of ex vivo ischemia time, freeze-thaw cycles, and cryopreservation time, the RIN of HCC tissues showed decreasing trends. Significant decreases in RIN of the tumor and normal tissues were observed at 6 and 2 hours of ex vivo ischemia time, respectively, and significantly decreased RIN of tumor tissues was observed after six freeze-thaw cycles and 6 years of cryopreservation. Conclusions: Preanalytical variables in the phase of preacquisition such as tumor grade, and in the postacquisition phase such as ex vivo ischemia time, freeze-thaw times, and freeze-storage time both have effects on RNA integrity of HCC tissues. Tissue-based translational research should pay attention to preanalytical variables when collecting and utilizing tumor tissues.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , ARN/genética , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Criopreservación , IsquemiaRESUMEN
Ferroptosis, as a new regulated mode of cell death, is mainly characterized by iron-dependent accumulation of lipid peroxides related to reactive oxygen species (ROS). However, the changes of peroxynitrite (ONOO-) during ferroptosis are not very clear due to the limited reports. On the other hand, ONOO- has become an endogenous toxicant leading to cell apoptosis and necrosis. Thus, it is urgent to develop molecular tools with high selectivity and sensitivity to monitor intracellular ONOO-. Herein, we presented a fluorescent probe, BTMO-PN, by introducing a phenylboronic acid ester as the ONOO- recognition site into benzothiazolyl derivative. BTMO-PN exhibited a rapid and marked fluorescence enhancement signal toward ONOO-, owing to the ONOO--triggered the cleavage of phenylboronic acid ester to release strongly fluorescent BTMO. Moreover, BTMO-PN could image endogenous and exogenous ONOO- changes in live cells. Importantly, using BTMO-PN, we demonstrated the up-generation of ONOO- levels in cancer cells during ferroptosis. Furthermore, BTMO-PN has successfully been applied for distinguishing tumor tissues from normal tissues with excellent contrast, making it great potential for cancer diagnosis by detecting the ONOO- changes.
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Ferroptosis , Neoplasias , Benzotiazoles , Ácidos Borónicos , Ésteres , Colorantes Fluorescentes , Humanos , Hierro , Peróxidos Lipídicos , Neoplasias/diagnóstico por imagen , Imagen Óptica , Ácido Peroxinitroso/metabolismo , Especies Reactivas de OxígenoRESUMEN
Distant metastasis is common in ocular uveal melanoma (uveal melanoma, UM) [1], with possible identification of relevant protein markers in peripheral blood [2], [3]. Proteomics analyses serve as a basis for the screening of new target proteins. However, it is difficult to determine whether the relevant proteins in peripheral blood are the same kinesins as those in primary lesions and metastases. Specially in this study, human UM cells (92.1) [4] were inoculated into the back of the eyeball and the brain of inbred line nude mice transplanted with enhanced green fluorescent protein (EGFP) [5], respectively, to simulate the growth of UM in situ and in brain metastases. A database was established as follows: Firstly, the xenograft was taken for monoclonal re-culture and amplification. Then, the cells after amplification (92.1-A in the back of the eyeball and 92.1-B in the brain) and their parent cells (92.1) were subjected to Tandem Mass Tag (TMT)-labeling proteomic analysis and liquid chromatography-mass spectrometry (LC-MS). Covering differential proteomes of three cell lines in a pairwise model, the data could be used to further screen the kinesins that play a vital role in regulating the growth of UM.
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The epithelial-mesenchymal transition (EMT) is an important process that drives progression, metastasis, and oncology treatment resistance in cancers. Also, the adjacent non-tumor tissue may affect the biological properties of cancers and have potential prognostic implications. Our study aimed to identify EMT-related genes in LGG samples, explore their impact on the biological properties of lower grade gliomas (LGG) through the multi-omics analysis, and reveal the potential mechanism by which adjacent non-tumor tissue participated in the malignant progression of LGG. Based on the 121 differentially expressed EMT-related genes between normal samples from the GTEx database and LGG samples in the TCGA cohort, we identified two subtypes and constructed EMTsig. Because of the genetic, epigenetic, and transcriptomic heterogeneity, malignant features including clinical traits, molecular traits, metabolism, anti-tumor immunity, and stemness features were different between samples with C1 and C2. In addition, EMTsig could also quantify the EMT levels, variation in prognosis, and oncology treatment sensitivity of LGG patients. Therefore, EMTsig could assist us in developing objective diagnostic tools and in optimizing therapeutic strategies for LGG patients. Notably, with the GSVA, we found that adjacent non-tumor tissue might participate in the progression, metastasis, and formation of the tumor microenvironment in LGG. Therefore, the potential prognostic implications of adjacent non-tumor tissue should be considered when performing clinical interventions for LGG patients. Overall, our study investigated and validated the effects of EMT-related genes on the biological properties from multiple perspectives, and provided new insights into the function of adjacent non-tumor tissue in the malignant progression of LGG.
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The development of effective biomedical technologies using magnetic nanoparticles (MNPs) for the tasks of oncotherapy and nanodiagnostics requires the development and implementation of new methods for the analysis of micro- and nanoscale distributions of MNPs in the volume of cells and tissues. The paper presents a new approach to three-dimensional analysis of MNP distributions - scanning magnetic force nanotomography as applied to the study of tumor tissues. Correlative reconstruction of MNP distributions and nanostructure features of the studied tissues made it possible to quantitatively estimate the parameters of three-dimensional distributions of composite nanoparticles based on silicon and iron oxide obtained by femtosecond laser ablation and injected intravenously and intratumorally into tumor tissue samples of B16/F1 mouse melanoma. The developed technology based on the principles of scanning probe nanotomography is applicable for studying the features of three-dimensional micro- and nanoscale distributions of magnetic nanoparticles in biomaterials, cells and tissues of various types.
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Nanopartículas de Magnetita , Melanoma Experimental , Nanopartículas , Animales , Materiales Biocompatibles , Fenómenos Magnéticos , Melanoma Experimental/diagnóstico por imagen , Ratones , Nanopartículas/químicaRESUMEN
Intratumoral environment as a hypoxic, non-inflamed "cold" state is difficult for many agents to accumulate and activate the immune system. Intrinsically, facultative anaerobic Salmonella VNP20009 target the tumor hypoxic areas, invade into tumor cells and exhibit an immune effect. Here we engineer the bacteria by decorating their surface with newly synthesized heptamethine cyanine dyes NHS-N782 and JQ-1 derivatives to obtain the biohybrid agent N-V-J, leading to the deep tumor targeted photothermal therapy and magnified immunotherapy. Due to the mitochondrial targeting capacity of NHS-N782, N-V-J becomes susceptive to the temperature rise when reaching tumors. This synergistic strategy promotes the systemic immunity by creating an inflamed "hot" tumor state from three different dimensions, which include the inherent immunogenicity of bacteria, the near-infrared laser triggered tumor antigens and the downregulation of PD-L1 expression. All these approaches result in effective and long-lasting T cell immune responses to prevent local and distant tumors for extended time. Leveraging the attenuated bacteria to transport dual drugs to the tumor tissues for self-synthetic vaccines provides a novel paradigm to enhance the bacteria-mediated cancer immunotherapy.
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Inmunoterapia , Neoplasias , Antígenos de Neoplasias , Bacterias , Línea Celular Tumoral , Humanos , Hipoxia , Inmunidad Celular , Inmunoterapia/métodos , Neoplasias/terapia , Fototerapia/métodos , Microambiente TumoralRESUMEN
Most human tumor tissues that are obtained for pathology and diagnostic purposes are formalin-fixed and paraffin-embedded (FFPE). To perform quantitative proteomics of FFPE samples, paraffin has to be removed and formalin-induced crosslinks have to be reversed prior to proteolytic digestion. A central component of almost all deparaffinization protocols is xylene, a toxic and highly flammable solvent that has been reported to negatively affect protein extraction and quantitative proteome analysis. Here, we present a 'green' xylene-free protocol for accelerated sample preparation of FFPE tissues based on paraffin-removal with hot water. Combined with tissue homogenization using disposable micropestles and a modified protein aggregation capture (PAC) digestion protocol, our workflow enables streamlined and reproducible quantitative proteomic profiling of FFPE tissue. Label-free quantitation of FFPE cores from human ductal breast carcinoma in situ (DCIS) xenografts with a volume of only 0.79 mm3 showed a high correlation between replicates (r2 = 0.992) with a median %CV of 16.9%. Importantly, this small volume is already compatible with tissue micro array (TMA) cores and core needle biopsies, while our results and its ease-of-use indicate that further downsizing is feasible. Finally, our FFPE workflow does not require costly equipment and can be established in every standard clinical laboratory.
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Parafina , Proteómica , Biopsia con Aguja Gruesa , Formaldehído , Humanos , Adhesión en Parafina , Proteoma/metabolismo , Proteómica/métodos , Fijación del TejidoRESUMEN
Owing to the important physiological sits of biothiols (Cys, Hcy, and GSH), developing accurate detection methods capable of qualitative and quantitative analysis of biothiols in living systems is needed for understanding the biological profile of biothiols. In this work, we have designed and synthesized a 4'-hydroxy-[1,1'-biphenyl]-4-carbonitrile modified with NBD group-based fluorescent probe, BPN-NBD, for sensitive detection of Cys/Hcy and GSH by dual emission signals via a single-wavelength excitation. BPN-NBD exhibited an obvious blue fluorescence (λmaxem = 475 nm) upon the treatment with GSH and reacted with Cys/Hcy to give a mixed blue-green fluorescence (λmaxem = 475 and 545 nm). Meanwhile, BPN-NDB performed sufficient selectivity, rapid detection (150 s), high sensitivity (0.011 µM for Cys, 0.015 µM for Hcy, and 0.003 µM for GSH) and could work via a single-wavelength excitation to analytes and had the ability to image Cys/Hcy from GSH in living MCF-7 cells, tumor tissues, and zebrafish by exhibiting different fluorescence signals. Overall, this work provided a powerful tool for thiols visualization in biological and medical applications.