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Nanotechnology has been widely utilized to enhance the intestinal absorption of natural bioactive compounds (BCs). However, the complicated and dynamic environment in the gastrointestinal tract (GIT) - characterized by digestive enzymes, mucus matrix, epithelial cell layer, and extensive metabolic activity - presents notable challenges to the oral delivery of BCs. To address these limitations, nanocarriers based on enzymatically hydrolyzed food protein polypeptides (EH-FPPs) have been developed, which exhibit high biocompatibility, loading capacity, and capacity to overcome different absorption barriers. This review provides a comprehensive summary of recent advancements in EH-FPP-assembled nanocarriers, including their self-assembly mechanisms, supramolecular structures, and loading approaches for natural BCs. Besides, it elucidates the how these nanocarriers enhance the intestinal absorption of BCs from the perspectives of controlled release, mucus penetration, transcytosis, and paracellular transport. Furthermore, it critically assesses the role of these nanocarriers in improving the overall bioavailability and therapeutic efficacy of encapsulated BCs, while also addressing their current limitations and future research directions. This review seeks to elucidate the complex relationships between the self-assembly process, multi-scale structure, and GIT fate of EH-FPP-based vehicles, which highlights their potential as advanced BC delivery systems for precision nutrition and individualized health.
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FMS-like tyrosine kinase 3 (FLT3) mutations are among the most common genetic lesions in acute myeloid leukemia (AML) and are frequently associated with high relapse rates and poor prognosis. Although current FLT3 inhibitors have improved outcomes, acquired resistance limits their long-term efficacy. Accordingly, developing new FLT3 inhibitors is important. We screened a natural-compound library to identify small molecules targeting the FLT3 protein. Kinase activity assays and cellular thermal shift assays (CETSA) collectively demonstrated the binding of canertinib to FLT3. Canertinib was tested in FLT3-mutated cells (32D, 32D-ITD, 32D-ITD+TKD, MV4-11 and blasts), and FLT3 wild-type cells. Cell viability was measured using the Cell Counting Kit-8 (CCK-8) assay; apoptosis was assessed by Annexin V staining; and phosphorylation of FLT3 and downstream pathways was analyzed by western blotting. A secondary-resistance model (MV4-11-ACR) was generated by treating MV4-11 cells with AC220. The effects of canertinib on cell viability, apoptosis, and FLT3 phosphorylation inhibition were evaluated in MV4-11-ACR cells. Canertinib directly targets the FLT3 protein. In FLT3-mutated expressed cells, canertinib effectively inhibited proliferation, induced apoptosis, and markedly reduced phosphorylation of FLT3 and its downstream effectors AKT, extracellular signal-regulated kinase (ERK), and STAT5, indicating precise on-pathway blockade of FLT3 signaling. Canertinib exerted similar effects on MV4-11-ACR cells as on the parental cells. Canertinib is an FLT3 inhibitor that directly targets the FLT3 protein to overcome FLT3 mutations and AC220 resistance in acute myeloid leukemia. These findings support canertinib as a promising candidate to overcome acquired resistance to FLT3 inhibitors.
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Antineoplásicos , Carbazóis , Leucemia Mieloide Aguda , Inibidores de Proteínas Quinases , Tirosina Quinase 3 Semelhante a fms , Tirosina Quinase 3 Semelhante a fms/antagonistas & inibidores , Tirosina Quinase 3 Semelhante a fms/genética , Tirosina Quinase 3 Semelhante a fms/metabolismo , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/enzimologia , Leucemia Mieloide Aguda/patologia , Humanos , Linhagem Celular Tumoral , Carbazóis/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Mutação , Antineoplásicos/farmacologia , MorfolinasRESUMO
PURPOSE: High programmed death-ligand 1 (PD-L1) expression is associated with unfavorable clinical outcomes in epidermal growth factor receptor (EGFR)-mutated lung adenocarcinomas (LUAD) patients treated with tyrosine kinase inhibitors (TKIs) or anti-PD-1/PD-L1 therapy, yet the underlying mechanisms are less explored. METHODS: Bulk RNA sequencing (RNA-seq) datasets were analyzed to investigate intertumoral transcriptional variations linked to PD-L1 expression. Immunohistochemistry (IHC) was utilized to quantify PD-L1 expression on tumor cells. Digital spatial profiling (DSP) was performed on 23 EGFR-mutated LUAD tissue samples to characterize transcriptomic differences in tumor cell (TC), immune cell (IM), and macrophage (MA) compartments between PD-L1 high and low groups. Furthermore, a publicly available DSP dataset was analyzed and IHC was conducted for validation. RESULTS: Analysis of RNA-seq datasets identified differentially expressed genes, signaling pathways, and immune profiles associated with PD-L1 expression. Compared to low PD-L1 tumors, high PD-L1 tumors exhibited increased infiltration of T regulatory cells (Tregs) and enhanced immunosuppressive signatures. DSP analysis revealed compartment-specific molecular disparities: TC segment in high PD-L1 tumors showed upregulated signatures of cell proliferation, invasion, and metastasis. IM segment displayed increased infiltration of immunosuppressive cells, including Tregs and myeloid-derived suppressor cells and upregulated expression of inhibitory immunomodulators CD276, HAVCR2, and LGALS9C. CONCLUSION: Combining bulk and spatial RNA-seq, this study characterized the molecular and immunological hallmarks of EGFR-mutated LUAD in the context of PD-L1 expression, providing new insights into the development of tailored therapeutic strategies for EGFR-mutated LUAD with high PD-L1 expression.
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Adenocarcinoma de Pulmão , Antígeno B7-H1 , Perfilamento da Expressão Gênica , Neoplasias Pulmonares , Mutação , Transcriptoma , Humanos , Antígeno B7-H1/metabolismo , Antígeno B7-H1/genética , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/patologia , Adenocarcinoma de Pulmão/imunologia , Receptores ErbB/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Mutação/genética , Regulação Neoplásica da Expressão Gênica , Transcriptoma/genética , Feminino , MasculinoRESUMO
FMS-like tyrosine kinase 3 (FLT3), a class III receptor tyrosine kinase, exhibits a mutation frequency of approximately 30% in acute myeloid leukemia (AML) patients and constitutes a critical therapeutic target. As representative type I and type II FLT3 inhibitors, respectively, gilteritinib and Quizartinib (AC220) are clinically employed in FLT3-mutant AML management. Gilteritinib inhibits both activated and inactivated FLT3 proteins, with additional inhibition of AXL targets to help overcome resistance. AC220 inhibits activated FLT3. The comet assay was systematically employed to quantify and compare DNA damage patterns induced by type I versus type II FLT3 inhibitors in mutant cells. The experimental results revealed that the DNA damage caused by AC220 was significantly higher than that caused by Gilteritinib. For strong DNA damage, FLT3 inhibitors can be combined with DNA damage repair inhibitors to target DNA repair defects. The results provide experimental support for the rational combination strategy of DNA damage-targeting drugs.
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Ensaio Cometa , Dano ao DNA , Inibidores de Proteínas Quinases , Tirosina Quinase 3 Semelhante a fms , Tirosina Quinase 3 Semelhante a fms/genética , Tirosina Quinase 3 Semelhante a fms/antagonistas & inibidores , Dano ao DNA/efeitos dos fármacos , Humanos , Ensaio Cometa/métodos , Benzotiazóis/farmacologia , Compostos de Fenilureia/farmacologia , Pirazinas/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Mutação , Linhagem Celular Tumoral , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/enzimologia , Compostos de AnilinaRESUMO
Aberrant lipid metabolism is intimately linked to tumor progression. As a pivotal post-translational modification, ubiquitination regulates diverse oncogenic processes. However, the interplay between ubiquitination and lipid metabolic dysregulation in pancreatic cancer (PC), along with its underlying molecular mechanisms, remains poorly understood. Here, it is demonstrated that glycolytic enzyme lactate dehydrogenase A (LDHA) potentiates lipid biosynthesis under the regulation of deubiquitinases. Specifically, PSMD14 directly binds and stabilizes LDHA through its deubiquitinase activity, resulting in intracellular lactate accumulation. Elevated lactate levels enhance histone lactylation marks, which transcriptionally activate ATP citrate lyase (ACLY) to promote malignant progression via fatty acid synthesis pathway activation. This study reveals a previously unrecognized role of PSMD14-derived lactate in mediating histone lactylation-coupled lipid deposition and tumor progression. Therapeutic co-targeting of PSMD14 and glycolytic lactylation significantly suppresses tumor growth in patient-derived xenograft models, suggesting a promising combinatorial strategy for pancreatic cancer treatment.
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ATP Citrato (pro-S)-Liase , Lactato Desidrogenase 5 , Neoplasias Pancreáticas , Complexo de Endopeptidases do Proteassoma , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Humanos , ATP Citrato (pro-S)-Liase/metabolismo , ATP Citrato (pro-S)-Liase/genética , Camundongos , Animais , Ubiquitinação , Linhagem Celular Tumoral , Progressão da Doença , Lactato Desidrogenase 5/metabolismo , Lactato Desidrogenase 5/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Complexo de Endopeptidases do Proteassoma/genética , Metabolismo dos Lipídeos/genética , Camundongos Nus , L-Lactato DesidrogenaseRESUMO
In this study, novel amphiphilic antioxidants, galloyl acylglycerols (GAGs), were successfully synthesized by integrating gallic acid (GA) with acylglycerols of varying acyl chain length (C12-C18) using mild Steglich esterification. The structural identities of these derivatives were confirmed through NMR and MS analysis. Further evaluation revealed that GAG-C18 exhibited excellent radical scavenging activity and superior antioxidant performance in both simple and complex liposomal dispersions compared to other gallic acid derivatives. Additionally, GAG-C18 effectively minimized the degradation and leakage of bioactives including curcumin and anthocyanins in multilayer and multivesicular liposomes. Microscopic investigation by confocal microscopy further demonstrated the visual results of the GAG-C18 to protect the liposome under oxidation. As an amphiphilic antioxidant, GAG-C18 demonstrated increased efficiency in lipid dispersions by preferentially accumulating in the oil-water interface, where oxidation predominantly occurs. The remarkable antioxidant performance of naturally derived amphiphilic GAG antioxidants makes them ideal candidates for ensuring oxidative stability in delivery systems.
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Antioxidantes , Ácido Gálico , Glicerol , Lipossomos , Lipossomos/química , Antioxidantes/química , Ácido Gálico/química , Oxirredução , Estrutura Molecular , Glicerol/química , Glicerol/análogos & derivadosRESUMO
FEN1 (Flap Endonuclease 1) is a key DNA nuclease involved in DNA replication and damage repair, playing a crucial role in maintaining genomic stability. Elevated levels of FEN1 activity are closely associated with the development of various cancers, making FEN1 a recognized biomarker and drug target for lung, breast, and prostate cancers. Conventional radioisotope tagging methods for detecting FEN1 nuclease activity present challenges in terms of convenience and safety. In this study, a DNA fragment substrate with a flap structure was synthesized in vitro. This substrate is labeled with a fluorophore at the 5' end of the flap, allowing the FEN1 nuclease to specifically recognize and cleave the designated site. FEN1 activity was then determined by detecting the fluorophore signal after gel electrophoresis. This method enables rapid and effective assessment of FEN1 nuclease activity and the screening of its inhibitors. It provides a sensitive and specific approach for analyzing FEN1 activity and evaluating the efficacy of small-molecule inhibitors. Furthermore, it contributes significantly to research on DNA damage repair and cancer development, offering a novel strategy for clinical research on disease mechanisms and the detection of genotoxic substances.
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Inibidores Enzimáticos , Endonucleases Flap , Bibliotecas de Moléculas Pequenas , Endonucleases Flap/antagonistas & inibidores , Endonucleases Flap/metabolismo , Endonucleases Flap/química , Endonucleases Flap/análise , Corantes Fluorescentes/química , Humanos , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/química , DNA/química , DNA/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologia , Bibliotecas de Moléculas Pequenas/químicaRESUMO
BACKGROUND: ERCC3, a crucial component of the nucleotide excision repair pathway, is implicated in the development and progression of various cancers and is a potential indicator of poor prognosis. However, the expression and function of ERCC3 in hepatocellular carcinoma (HCC) remain unclear. This study aimed to investigate the expression of ERCC3 in HCC tissues and its clinical significance, focusing on elucidating its potential mechanisms and therapeutic value in immunotherapy. METHODS: The differential expression and genetic variation characteristics of ERCC3 across various cancers were evaluated using the TCGA database. The expression and prognostic value of ERCC3 in HCC were analyzed by integrating TCGA, GEO, and ICGC datasets. Independent prognostic value of ERCC3 expression levels in HCC was assessed using Cox regression analysis, Kaplan-Meier survival analysis, receiver operating characteristic curves, and nomograms. Pathway association scores were determined using ssGSEA to reveal the biological functions of ERCC3 in HCC and its potential clinical efficacy in immunotherapy. Stable transient cell lines were established by infecting HepG2 cells with lentivirus overexpressing ERCC3. The effects of ERCC3 on HCC cell biological phenotypes were evaluated using RTCA, wound healing, and Transwell assays. Cell cycle distribution and apoptosis were detected by flow cytometry. Transcriptome sequencing was performed to explore the impact of ERCC3 overexpression on the expression of signaling pathway-related genes in HCC. RESULTS: The study revealed that ERCC3 is aberrantly expressed in various tumors, with significantly higher mRNA and protein levels in HCC tissues compared to normal tissues. High ERCC3 expression was significantly correlated with poor survival outcomes in HCC patients. Multivariate Cox regression analysis revealed that ERCC3 expression level is an independent prognostic factor for overall survival (P = 0.014). Gene sets associated with the high ERCC3 group were significantly involved in multiple immune pathways and tumor progression-related pathways, and ERCC3 expression was significantly correlated with immune checkpoints in HCC. Overexpression of ERCC3 promoted the proliferation and migration of HCC cells and influenced cell cycle progression. Transcriptome sequencing analysis indicated that ERCC3 overexpression regulated the proliferation of HCC cells, participated in multiple pro-inflammatory pathways, induced the formation of an inflammatory tumor microenvironment, and promoted HCC progression. CONCLUSION: This study is the first to reveal the association between high ERCC3 expression and poor prognosis in HCC and to elucidate its immunomodulatory role in HCC. Unlike previous studies, we found that ERCC3 promotes HCC progression by regulating the inflammatory microenvironment and immune checkpoints. These findings establish a novel theoretical foundation for the development of targeted immunotherapies for HCC and provide new insights into the molecular mechanisms underlying ERCC3's role in HCC.
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BACKGROUND: Dysregulation or abnormality of the programmed cell death (PCD) pathway is closely related to the occurrence and development of many tumors, including acute myeloid leukemia (AML). Studying the abnormal characteristics of PCD pathway-related molecular markers can provide a basis for prognosis prediction and targeted drug design in AML patients. METHODS: A total of 1394 genes representing 13 different PCD pathways were examined in AML patients and healthy donors. The upregulated genes were analyzed for their ability to predict overall survival (OS) individually, and these prognostic genes were subsequently combined to construct a PCD-related prognostic signature via an integrated approach consisting of 101 models based on ten machine learning algorithms. RNA transcriptome and clinical data from multiple AML cohorts (TCGA-AML, GSE106291, GSE146173 and Beat AML) were obtained to develop and validate the AML prognostic model. RESULTS: A total of 214 upregulated PCD-related genes were identified in AML patients, 39 of which were proven to be prognostic genes in the training cohort. On the basis of the average C-index and number of model genes identified from the machine learning combinations, a PCD index was developed and validated for predicting AML OS. A prognostic nomogram was then generated and validated on the basis of the PCD index, age and ELN risk stratification in the Beat AML cohort and the GSE146173 cohort, revealing satisfactory predictive power (AUC values ≥ 0.7). With different mutation patterns, a higher PCD index was associated with a worse OS. The PCD index was significantly related to higher scores for immunosuppressive cells and mature leukemia cell subtypes. As the gene most closely related to the PCD index, the expression of SMAD3 was further validated in vitro. AML cells harboring KMT2A rearrangements were more sensitive to the SMAD3 inhibitor SIS3, and the expression of the autophagy-related molecular marker LC3 was increased in KMT2A-rearranged cell lines after SIS3 monotherapy and combined treatment. CONCLUSION: The PCD index and SMAD3 gene expression levels have potential prognostic value and can be used in targeted therapy for AML, and these findings can lead to the development of effective strategies for the combined treatment of high-risk AML patients.
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Biomarcadores Tumorais , Leucemia Mieloide Aguda , Aprendizado de Máquina , Proteína Smad3 , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/mortalidade , Leucemia Mieloide Aguda/patologia , Leucemia Mieloide Aguda/tratamento farmacológico , Prognóstico , Proteína Smad3/genética , Proteína Smad3/antagonistas & inibidores , Proteína Smad3/metabolismo , Masculino , Feminino , Biomarcadores Tumorais/genética , Perfilamento da Expressão Gênica , Pessoa de Meia-Idade , Apoptose/genética , Transcriptoma , NomogramasRESUMO
This review summarizes the applications and research progress of organoid models in colorectal cancer research. First, the high incidence and mortality rates of colorectal cancer are introduced, emphasizing the importance of organoids as a research model. Second, this review provides a detailed introduction to the concept, biological properties, and applications of organoids, including their strengths in mimicking the structural and functional aspects of organs. This article further analyzes the applications of adult stem cell-derived and pluripotent stem cell-derived organoids in colorectal cancer research and discusses advancements in organoids for basic research, drug research and development, personalized treatment evaluation and prediction, and regenerative medicine. Finally, this review summarizes the prospects for applying organoid technology in colorectal cancer research, emphasizing its significant value in improving patient survival rates. In conclusion, this review systematically explains the applications of organoids in colorectal cancer research, highlighting their tremendous potential and promising prospects in basic research, drug research and development, personalized treatment evaluation and prediction, and regenerative medicine.
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Oral targeted delivery systems for food bioactive compounds have attracted considerable attention for improving the efficiency of nutritional interventions. In this study, a hierarchical curcumin carrier with sequence-targeting capability was fabricated to mitigate colitis through a convenient and environmentally friendly approach. Initially, curcumin-loaded nanostructured lipid carriers (NLC) with hyaluronic acid (HA) moieties (HA-NLC) were prepared using ovalbumin-HA conjugates as emulsifiers. Subsequently, HA-NLC was immobilized within a calcium pectinate matrix using electrospray, leading to the formation of supramolecular microspheres (HA-NLC@MPs) approximately 140 µm in size. Results suggested that the pectin matrix preserved the integrity of the carrier and prevented curcumin leakage during transit through the upper gastrointestinal tract, while selectively degrading in response to colon bacteria. Moreover, the exposed HA moieties on the released HA-NLC facilitated the transcellular absorption of curcumin by inflamed colonic enterocytes through cluster of differentiation-44 receptor-mediated endocytosis. In vitro and in vivo studies demonstrated predominant curcumin accumulation in the colorectal tissues of colitis mice using HA-NLC@MPs as carrier. Curcumin delivered via HA-NLC@MPs demonstrated superior effects in alleviating colitis compared with that in the nanocarriers, through the modulation of macrophage polarization, TLR4/MyD88/NF-κB signaling cascade, and gut microbiota homeostasis.
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Anti-Inflamatórios , Colite , Curcumina , Microesferas , Curcumina/administração & dosagem , Suplementos Nutricionais , Sistemas de Liberação de Medicamentos , Suco Gástrico/química , Suco Gástrico/metabolismo , Animais , Ratos , Fezes/química , Lipídeos/química , Células CACO-2 , Humanos , Linhagem Celular Tumoral , Camundongos , Colite/induzido quimicamente , Colite/tratamento farmacológico , Distribuição Aleatória , Microbioma Gastrointestinal , Pectinas/química , Anti-Inflamatórios/administração & dosagem , Estresse Oxidativo/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
Cancer immunotherapy has attracted great attention as a potential therapeutic approach for advanced malignancies due to its promising survival benefits. Comprehension of intricate interactions between the tumor microenvironment (TME) and immune checkpoint inhibitors (ICIs) is crucial for optimizing and improving immunotherapies. Currently, several experimental strategies are available to monitor this complexity but most of them fail to facilitate real-time monitoring of the immune response such as cellular phagocytosis and cytolysis. Consequently, the application of intravital imaging has been extensively studied in the domain of cancer immunotherapy. Intravital imaging has been proven to be a powerful real-time imaging modality that provides insights into intratumoral immune responses, cellular metabolic signatures, tumor vasculature, and cellular functions. This review aims to provide a comprehensive overview of the latest research on intravital imaging in cancer immunotherapy, especially addressing how intravital imaging sheds light on essential features of tumor immunity, immune infiltrations, tumor angiogenesis, and aids in the clarification of underlying immunotherapeutic mechanisms. Moreover, a variety of labeling tools, imaging windows and models for real-time visualizations of TME are also summarized. We will also investigate the full potential of using intravital imaging to circumvent the limitations of currently available imaging modalities, which hold promise to advent efficient immunotherapy for cancer patients.
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Imunoterapia , Microscopia Intravital , Neoplasias , Humanos , Imunoterapia/métodos , Neoplasias/terapia , Neoplasias/imunologia , Neoplasias/diagnóstico por imagem , Microambiente Tumoral/imunologia , Animais , Microscopia Intravital/métodosRESUMO
The substances that cause abnormal DNA structures are known as DNA damage-inducing factors, and their resulting DNA damage has been extensively studied and proven to be closely related to cancer, neurodegenerative diseases, and aging. Prolonged exposure to DNA damage-inducing factors can lead to a variety of difficult-to-treat diseases, yet these factors have not been well summarized. It is crucial to use a combination of environmental science and life science to gain a deep understanding of the environmental sources and biological consequences of DNA damage-inducing factors for mechanistic research and prevention of diseases such as cancer. This article selected 14 representative carcinogenic exogenous DNA damage-inducing factors and summarized them through a literature search, including both exogenous and endogenous DNA damage factors, and explored the types of DNA damage caused by the relevant damage factors.
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Acetyl CoA acetyltransferase 1 (ACAT1), a mitochondrial enzyme, is mainly involved in the formation and decomposition of ketones, isoleucine, and fatty acids. Previous clinical studies showed that mutations in the ACAT1 gene lead to ketoacidosis, Notably the role of ACAT1 in human cancer' pathogenesis varies depending on cancer type, and its specific role in gastric cancer remains largely unknown. In the current study, we found that the expression of ACAT1 in primary late-stage gastric cancer tumor tissues was significantly lower than in early-stage tumors. This observation was further confirmed in high-grade gastric cancer cell line MKN45. The expression of CD44 and OCT4 was decreased, while CD24 expression was increased by overexpressing ACAT1 in MKN45 gastric cancer cells. Moreover, the ability of gastric cancer cells to form colonies on soft agar was also reduced by ACAT1 overexpression. Likewise, overexpression of ACAT1 inhibited epithelial mesenchymal transition (EMT) in gastric cancer cells evidenced by increased expression of the epithelial marker E-Cadherin, decreased expression of mesenchymal marker vimentin, and decreased expression levels of SNAI 1/3. In addition, ACAT1 overexpression inhibited cell migration and invasion, improved the response to 5-Fluorouracil (5-FU) and etoposide. In contrast, inhibition of ACAT1 activity promoted the proliferation of gastric cancer cells. The xenotransplantation results in nude mice showed that overexpression of ACAT1 in gastric cancer cells inhibited tumor growth in vivo. In addition, the low expression of ACAT1 in gastric cancer was further validated by searching public databases and conducting bioinformatic analyses. Mechanistically, bioinformatic analysis found that the inhibitory effect of ACAT1 in gastric cancer may be related to the Adipocytokine Signaling Pathway, Ppar Signaling Pathway, Propanoate Metabolism and P53 Signaling Pathway. Correlation analysis indicated ACAT1 mRNA expression was correlated with immune infiltrates. Collectively, our data show that ACAT1 induces pronounced inhibitory effects on gastric cancer initiation and development, which may impact future strategies to treat this aggressive cancer.
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Acetil-CoA C-Acetiltransferase , Transição Epitelial-Mesenquimal , Mitocôndrias , Neoplasias Gástricas , Neoplasias Gástricas/patologia , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Humanos , Animais , Camundongos , Linhagem Celular Tumoral , Acetil-CoA C-Acetiltransferase/metabolismo , Acetil-CoA C-Acetiltransferase/genética , Mitocôndrias/metabolismo , Regulação Neoplásica da Expressão Gênica , Camundongos Nus , Movimento Celular , Proliferação de Células , Feminino , Masculino , Ensaios Antitumorais Modelo de Xenoenxerto , Fluoruracila/farmacologiaRESUMO
8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) base is the predominant form of commonly observed DNA oxidative damage. DNA impairment profoundly impacts gene expression and serves as a pivotal factor in stimulating neurodegenerative disorders, cancer, and aging. Therefore, precise quantification of 8-oxoG has clinical significance in the investigation of DNA damage detection methodologies. However, at present, the existing approaches for 8-oxoG detection pose challenges in terms of convenience, expediency, affordability, and heightened sensitivity. We employed the sandwich enzyme-linked immunosorbent assay (ELISA) technique, a highly efficient and swift colorimetric method, to detect variations in 8-oxo-dG content in MCF-7 cell samples stimulated with different concentrations of hydrogen peroxide (H2O2). We determined the concentration of H2O2 that induced oxidative damage in MCF-7 cells by detecting its IC50 value in MCF-7 cells. Subsequently, we treated MCF-7 cells with 0, 0.25, and 0.75 mM H2O2 for 12 h and extracted 8-oxo-dG from the cells. Finally, the samples were subjected to ELISA. Following a series of steps, including plate spreading, washing, incubation, color development, termination of the reaction, and data collection, we successfully detected changes in the 8-oxo-dG content in MCF-7 cells induced by H2O2. Through such endeavors, we aim to establish a method to evaluate the degree of DNA oxidative damage within cell samples and, in doing so, advance the development of more expedient and convenient approaches for DNA damage detection. This endeavor is poised to make a meaningful contribution to the exploration of associative analyses between DNA oxidative damage and various domains, including clinical research on diseases and the detection of toxic substances.
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8-Hidroxi-2'-Desoxiguanosina , Dano ao DNA , Ensaio de Imunoadsorção Enzimática , Peróxido de Hidrogênio , Estresse Oxidativo , Humanos , Dano ao DNA/efeitos dos fármacos , Células MCF-7 , Ensaio de Imunoadsorção Enzimática/métodos , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análiseRESUMO
BACKGROUND: Hepatocellular carcinoma (HCC) has high morbidity and mortality worldwide. Excision repair cross-complement 3 (ERCC3), a key functional gene in the nucleotide excision repair (NER) pathway, is commonly mutated or overexpressed in cancers and is thought to be a key gene contributing to the development of HCC. The characteristics of immune cell infiltration in the global tumor microenvironment (TME) mediated by ERCC3 and its related key genes in HCC are still unclear. The aim of this study was to integrate the role of ERCC3-related key genes in assessing the TME cell infiltration characteristics, immunotherapy efficacy, and prognosis of HCC patients. This study provides a theoretical basis for the study of immunological mechanisms and prognosis prediction in HCC. METHODS: The HCC cohort from the TCGA database included 50 normal samples and 374 tumor samples to compare the differences in ERCC3-related gene expression and prognosis between liver tumor tissues and normal liver tissues and to analyze the extent to which different genes infiltrated TME cells by quantifying the relative abundance of 24 cells through single-sample genome enrichment analysis (ssGSEA). A risk score associated with the ERCC3 gene was constructed using the least absolute shrinkage and selection operator (LASSO) Cox regression model. RESULTS: The expression of 11 ERCC3-related genes was significantly upregulated in HCC tumor tissues compared to normal liver tissues, and high expression of these genes was significantly associated with poor prognosis in HCC patients. The key genes (11 ERCC3-related genes) were closely associated with the nucleic acid reduction signaling pathway in nucleic acid metabolism and the viral oncogenic pathway, suggesting that these key genes may play a role in tumor cell proliferation, migration, and invasion, as well as in the pathogenesis of virus-associated HCC. In addition, the infiltration characteristics of TME immune cells in normal and tumor tissues were different. Immune and mesenchymal activity was significantly lower in tumor tissues than in healthy liver tissues. This study revealed that key genes were significantly positively correlated with CTLA4 and enriched in central memory CD4 T cells, effector memory CD4 T cells, activated CD4 T cells, and type 2 T helper cells. The prognostic model constructed by regression analysis could better distinguish patients into high-risk and low-risk groups, and the survival analysis showed that the survival time of patients with high-risk score subtypes was significantly lower than that of patients with low-risk scores and that the high-risk group contained higher levels of immune-suppressive cells, which may be a mediator of immune escape. Moreover, multivariate analyses showed that the risk score profile is a reliable and unbiased biomarker for assessing the prognosis of HCC patients, and its value in predicting the outcome of immunotherapy was also confirmed. CONCLUSION: This study revealed a novel genetic signature that is significantly associated with TME cell infiltration and prognosis in HCC patients. It demonstrated that the combined action of multiple key genes associated with ERCC3 plays a crucial role in shaping the diversity and complexity of TME cell infiltrates. Evaluating the combined characteristics of multiple key genes associated with ERCC3 can help predict the outcome of immunotherapy in patients and provide new potential targets for immuno-individualized therapeutic studies on HCC.
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Conventional radical grafting of proteins with catechins consumed the most antioxidant-active hydroxyls during grafting, thus failing to effectively retain antioxidant activity in conjugates. In this study, a novel strategy of selective protection of the most reactive hydroxyls before grafting was developed to preserve the most reactive hydroxyls and effectively retain antioxidant activity in conjugates. Selective protection of the most reactive hydroxyls of (-)-epigallocatechin-3-gallate (EGCG) was successfully realized in a yield of 87% applying trimethyl orthopropionate and catalytic calcium triflate at 40 °C. The novel ovalbumin (OVA)-EGCG conjugate with 93% grafting ratio was prepared by radical grafting with the selectively protected EGCG and subsequent deprotection. Substantially enhanced antioxidant performance of the novel OVA-EGCG conjugate in liposomes was unveiled with notably reduced curcumin degradation and leakage. The strategy and approaches developed in this study will be valuable to effectively improve the antioxidant activities of protein-catechin grafting conjugates.
Assuntos
Antioxidantes , Catequina , Ovalbumina , Ovalbumina/química , Catequina/química , Catequina/análogos & derivados , Antioxidantes/química , Lipossomos/químicaRESUMO
PURPOSE: Pcancreatic cancer (PC) is a common tumor of the digestive tract with an insidious onset and high malignancy potential. Currently, surgery is the only effective treatment modality. Therefore, it is crucial to discover new targeted therapeutic modalities. We studied whether transgelin 2 (TAGLN2) targeted control of actin-related protein 2/3 complex subunit 5 (ARPC5)-mediated activation of the MEK/ERK signaling pathway to Influences the proliferation, invasion, and metastasis of pancreatic cancer cells. METHODS: The effects of TAGLN2 overexpression and knockdown on the proliferative viability and invasive metastatic ability of pancreatic cancer cells were verified through in vitro and in vivo assays via constructing a stable lentiviral transfection of human pancreatic cancer cell lines PANC-1 and SW1990. Bioinformatics analysis was used to predict the relationship between TAGLN2 and ARPC5. These findings were subsequently verified through protein profiling, immunofluorescence (IF), and coimmunoprecipitation (CO-IP) assays. In vitro experiments were also conducted to confirm the effect of TAGLN2 modulation on ARPC5 expression, which subsequently affects the proliferation and invasive metastatic ability of pancreatic cancer cells. The study analyzed the relationship between TAGLN2 and the MEK/ERK signaling pathway through bioinformatics and in vitro experiments with the MEK signaling pathway inhibitor U0126. RESULTS: TAGLN2 is expressed at high levels in pancreatic cancer cell lines, and its expression is positively correlated with poor prognosis of pancreatic cancer. ARPC5 is a direct target of TAGLN2 and is associated with the MEK/ERK signaling pathway. In vivo and ex vivo experiments confirmed that overexpression of TAGLN2 promoted the proliferation, invasion, and metastasis of pancreatic cancer cells, and silencing ARPC5 reversed these effect. CONCLUSION: Our research revealed that TAGLN2 protein binds to ARPC5 protein and contributes to increased ARPC5 expression and activation of the MEK/ERK signaling pathway. This activation promotes pancreatic cancer cell growth, infiltration, and spread. Hence, TAGLN2 is a potential viable therapeutic target in pancreatic cancer and represents a novel therapeutic approach.
Assuntos
Proliferação de Células , Sistema de Sinalização das MAP Quinases , Proteínas dos Microfilamentos , Proteínas Musculares , Invasividade Neoplásica , Neoplasias Pancreáticas , Animais , Humanos , Camundongos , Linhagem Celular Tumoral , Movimento Celular , Regulação Neoplásica da Expressão Gênica , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas dos Microfilamentos/metabolismo , Proteínas dos Microfilamentos/genética , Proteínas Musculares/metabolismo , Proteínas Musculares/genética , Metástase Neoplásica , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/genéticaRESUMO
Programmed cell death (PCD) is a basic process of life that is closely related to the growth, development, aging and disease of organisms and is one of the hotspots of life science research today. PCD is a kind of genetic control, autonomous and orderly important cell death that involves the activation, expression, and regulation of a series of genes. In recent years, with the deepening of research in this field, new mechanisms of multiple PCD pathways have been revealed. This article reviews and summarizes the multiple PCD pathways that have been discovered, analyses and compares the morphological characteristics and biomarkers of different types of PCD, and briefly discusses the role of various types of PCD in the diagnosis and treatment of different diseases, especially malignant tumors.