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Marine Drugs Oct 2016An extract prepared from the culture of a marine-derived actinomycete sp. ZZ338 was found to have significant antimicrobial and antiproliferative activities. A chemical...
An extract prepared from the culture of a marine-derived actinomycete sp. ZZ338 was found to have significant antimicrobial and antiproliferative activities. A chemical investigation of this active extract resulted in the isolation of three known bioactive actinomycins (-) and two new metabolites ( and ). The structures of the isolated compounds were identified as actinomycins D (), V (), X (), 2-acetylamino-3-hydroxyl-4-methyl-benzoic acid methyl ester (), and -1-(4-methylaminophenylmethyl)-2-oxo-propyl acetamide () based on their nuclear magnetic resonance (NMR) and high resolution electrospray ionization mass spectroscopy (HRESIMS) data as well as their optical rotation. This class of new compound had never before been found from a natural resource. Three known actinomycins showed activities in inhibiting the proliferation of glioma cells and the growth of methicillin-resistant , , and and are responsible for the activity of the crude extract. Actinomycin D () was also found to downregulate several glioma metabolic enzymes of glycolysis, glutaminolysis, and lipogenesis, suggesting that targeting multiple tumor metabolic regulators might be a new anti-glioma mechanism of actinomycin D. This is the first report of such a possible mechanism for the class of actinomycins.
Topics: Animals; Anti-Infective Agents; Antineoplastic Agents; Candida albicans; Dactinomycin; Drug Screening Assays, Antitumor; Escherichia coli; Humans; Inhibitory Concentration 50; Oceans and Seas; Rats; Staphylococcus aureus; Streptomyces; Structure-Activity Relationship; Tumor Cells, Cultured
PubMed: 27727167
DOI: 10.3390/md14100181 -
Marine Drugs Oct 2021Actinomycin (Act) V, an analogue of Act D, presented stronger antitumor activity and less hepatorenal toxicity than Act D in our previous studies, which is worthy of...
Actinomycin (Act) V, an analogue of Act D, presented stronger antitumor activity and less hepatorenal toxicity than Act D in our previous studies, which is worthy of further investigation. We hereby report that Act V induces apoptosis via mitochondrial and PI3K/AKT pathways in colorectal cancer (CRC) cells. Act V-induced apoptosis was characterized by mitochondrial dysfunction, with loss of mitochondria membrane potential (MMP) and cytochrome c release, which then activated cleaved caspase-9, cleaved caspase-3, and cleaved PARP, revealing that it was related to the mitochondrial pathway, and the apoptotic trendency can be reversed by caspase inhibitor Z-VAD-FMK. Furthermore, we proved that Act V significantly inhibited PI3K/AKT signalling in HCT-116 cells using cell experiments in vitro, and it also presented a potential targeted PI3Kα inhibition using computer docking models. Further elucidation revealed that it exhibited a 28-fold greater potency than the PI3K inhibitor LY294002 on PI3K inhibition efficacy. Taken together, Act V, as a superior potential replacement of Act D, is a potential candidate for inhibiting the PI3K/AKT pathway and is worthy of more pre-clinical studies in the therapy of CRC.
Topics: Animals; Antineoplastic Agents; Apoptosis; Aquatic Organisms; Cell Line, Tumor; Dactinomycin; Humans; Membrane Potential, Mitochondrial; Mitochondria; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Streptomyces
PubMed: 34822470
DOI: 10.3390/md19110599 -
International Journal of Molecular... Nov 2021() is a common pathogen that causes various serious diseases, including chronic infections. Discovering new antibacterial agents is an important aspect of the...
() is a common pathogen that causes various serious diseases, including chronic infections. Discovering new antibacterial agents is an important aspect of the pharmaceutical field because of the lack of effective antibacterial drugs. In our research, we found that one anti- substance is actinomycin D, originating from (); then, we further focused on the anti- ability and the omics profile of in response to actinomycin D. The results revealed that actinomycin D had a significant inhibitory activity on with a minimum inhibitory concentration (MIC) of 2 μg/mL and a minimum bactericidal concentration (MBC) of 64 μg/mL. Bacterial reactive oxygen species (ROS) increased 3.5-fold upon treatment with actinomycin D, as was measured with the oxidation-sensitive fluorescent probe DCFH-DA, and HO increased 3.5 times with treatment by actinomycin D. Proteomics and metabolomics, respectively, identified differentially expressed proteins in control and treatment groups, and the co-mapped correlation network of proteomics and metabolomics annotated five major pathways that were potentially related to disrupting the energy metabolism and oxidative stress of . All findings contributed to providing new insight into the mechanisms of the anti- effects of actinomycin D originating from
Topics: Anti-Bacterial Agents; Dactinomycin; Metabolomics; Proteomics; Reactive Oxygen Species; Staphylococcus aureus; Streptomyces
PubMed: 34830114
DOI: 10.3390/ijms222212231 -
Disease Markers 2022Macrophages are heterogeneous cells that can be polarized into M1 or M2 phenotype. mA "reader" YTH domain family protein 2 (YTHDF2) has been the mA binding protein with...
Macrophages are heterogeneous cells that can be polarized into M1 or M2 phenotype. mA "reader" YTH domain family protein 2 (YTHDF2) has been the mA binding protein with the highest activity, which can recognize and disturb mA-containing mRNA in processing bodies to reduce mRNA stability. YTHDF2 is recently identified as an effective RNA binding protein that modulates inflammatory gene levels within inflammatory responses. However, the role of YTHDF2 in M1/M2 macrophage polarization has not been reported. We established a M1/M2 macrophage polarization model using bone-marrow-derived macrophages and found that the expression levels of YTHDF2 in M1/M2 macrophages were both elevated. YTHDF2-knockdown macrophage polarization model was then established, and through qPCR, ELISA, and FACS, we discovered that suppressing YTHDF2 encouraged M1 polarization but restrained M2 polarization. In M1 macrophages, YTHDF2 silencing had no significant effect on p53 expression; however, in YTHDF2 knockdown, M2 macrophage p53 expression was remarkably upregulated. p53 inhibitor PFT- was then applied and revealed that suppressing p53 simultaneously promoted YTHDF2-silenced M1 polarization and facilitated M2 macrophage polarization. Actinomycin D assays were further utilized to examine the mRNA degradation level of different cytokines, and p53 mRNA degradation in YTHDF2-depleted M2 cells was discovered impeded. Western Blot analysis also implied that a deficit in YTHDF2 expression may activate MAPK and NF-B pathways. In this study, YTHDF2 induces M2 macrophage polarization by promoting the degradation of p53 mRNA. YTHDF2 suppresses M1 macrophage polarization by inhibiting NF-B, p38, and JNK signaling pathways, yet p53 remains unaffected in YTHDF2-silenced M1 macrophages.
Topics: NF-kappa B; Tumor Suppressor Protein p53; Dactinomycin; Signal Transduction; Macrophages; Cytokines; RNA-Binding Proteins; RNA, Messenger
PubMed: 36277978
DOI: 10.1155/2022/3153362 -
Nucleic Acids Research May 2023Combination cancer chemotherapy is one of the most useful treatment methods to achieve a synergistic effect and reduce the toxicity of dosing with a single drug. Here,...
Combination cancer chemotherapy is one of the most useful treatment methods to achieve a synergistic effect and reduce the toxicity of dosing with a single drug. Here, we use a combination of two well-established anticancer DNA intercalators, actinomycin D (ActD) and echinomycin (Echi), to screen their binding capabilities with DNA duplexes containing different mismatches embedded within Watson-Crick base-pairs. We have found that combining ActD and Echi preferentially stabilised thymine-related T:T mismatches. The enhanced stability of the DNA duplex-drug complexes is mainly due to the cooperative binding of the two drugs to the mismatch duplex, with many stacking interactions between the two different drug molecules. Since the repair of thymine-related mismatches is less efficient in mismatch repair (MMR)-deficient cancer cells, we have also demonstrated that the combination of ActD and Echi exhibits enhanced synergistic effects against MMR-deficient HCT116 cells and synergy is maintained in a MMR-related MLH1 gene knockdown in SW620 cells. We further accessed the clinical potential of the two-drug combination approach with a xenograft mouse model of a colorectal MMR-deficient cancer, which has resulted in a significant synergistic anti-tumour effect. The current study provides a novel approach for the development of combination chemotherapy for the treatment of cancers related to DNA-mismatches.
Topics: Humans; Animals; Mice; Dactinomycin; Echinomycin; Thymine; Base Sequence; Binding Sites; Nucleic Acid Conformation; DNA; Colorectal Neoplasms
PubMed: 36919604
DOI: 10.1093/nar/gkad156 -
Journal of Virology Jun 1968Deoxyribonucleic acid (DNA) extracted from phage SPP1 is highly infectious on Bacillus subtilis competent cells; the efficiency of infection is 5 x 10(3) to 6 x 10(3)...
Deoxyribonucleic acid (DNA) extracted from phage SPP1 is highly infectious on Bacillus subtilis competent cells; the efficiency of infection is 5 x 10(3) to 6 x 10(3) phage equivalents per plaque-forming unit. This DNA was used to study the relationship between competence for transfection and for transformation. The experiments were concerned with the frequency of infection and transformation in mutants exhibiting different levels of competence, the effect of periodate on competence for infection and for transformation, the competition between phage and bacterial DNA, the transformation of cells preinfected with phage DNA, and the infection of cells pretreated with bacterial DNA. The data show that B. subtilis cells competent for transformation are also competent for transfection and vice versa; transfection with phage DNA represents, therefore, a simple way to measure the total number of competent cells in a culture. The fraction of competent cells, determined by SPP1 DNA infection, varied from 10(-2) to 7 x 10(-2).
Topics: Bacillus subtilis; Bacteriophages; Centrifugation, Density Gradient; DNA, Bacterial; DNA, Viral; Dactinomycin; Drug Resistance, Microbial; Genetics, Microbial; Kinetics; Periodic Acid; Transformation, Genetic
PubMed: 4177104
DOI: 10.1128/JVI.2.6.587-593.1968 -
Scientific Reports Jun 2017Neo-actinomycins A and B (1 and 2), two new natural actinomycins featuring an unprecedented tetracyclic 5H-oxazolo[4,5-b]phenoxazine chromophore, were isolated from the...
Neo-actinomycins A and B (1 and 2), two new natural actinomycins featuring an unprecedented tetracyclic 5H-oxazolo[4,5-b]phenoxazine chromophore, were isolated from the marine-derived actinomycete Streptomyces sp. IMB094. Their structures were elucidated by spectroscopic analyses. The presence of this ring system was proposed to originate from a condensation between actinomycin D (3) with α-ketoglutarate and pyruvate, respectively. Compound 1 showed potent cytotoxic activities against human cancer HCT116 and A549 cell lines in the nanomolar range (IC: 38.7 and 65.8 nM, respectively) and moderate antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) strains.
Topics: Anti-Bacterial Agents; Antibiotics, Antineoplastic; Aquatic Organisms; Biological Products; Cell Line, Tumor; Cell Survival; Dactinomycin; Enterococcus; Humans; Inhibitory Concentration 50; Molecular Structure; Spectrum Analysis; Staphylococcus aureus; Streptomyces
PubMed: 28620204
DOI: 10.1038/s41598-017-03769-8 -
Methods in Enzymology 2009The myosin superfamily consists of more than 35 classes (each consisting of multiple isoforms) that have diverse cellular activities. The reaction pathway of the...
The myosin superfamily consists of more than 35 classes (each consisting of multiple isoforms) that have diverse cellular activities. The reaction pathway of the actin-activated myosin ATPase appears to be conserved for all myosin isoforms, but the rate and equilibrium constants that define the ATPase pathway vary significantly across the myosin superfamily, resulting in kinetic differences that that allow myosins to carry out diverse mechanical functions. Therefore, it is important to determine the lifetimes and relative populations of the key biochemical intermediates to obtain an understanding of a particular myosin's cellular function. This chapter provides procedures for determining the overall and individual rate and equilibrium constants of the actomyosin ATPase cycle, including actomyosin binding and dissociation, ATP binding, ATP hydrolysis, phosphate release, and ADP release and binding. Many of the methods described in the chapter are applicable to the characterization of other ATPase enzymes.
Topics: Animals; Computer Simulation; Dactinomycin; Enzyme Stability; Humans; Kinetics; Myosins; Protein Binding; Thermodynamics
PubMed: 19289206
DOI: 10.1016/S0076-6879(08)04206-7 -
Apoptosis : An International Journal on... Jun 2022Actinomycin D (ActD) was the first anticancer antibiotic approved for the management of human cancers. However, the notorious toxicity profile limits its widespread...
Actinomycin D (ActD) was the first anticancer antibiotic approved for the management of human cancers. However, the notorious toxicity profile limits its widespread application in cancers, including cancers of the aerodigestive tract. Recent studies show that combining low-dose ActD with existing chemotherapies could potentially protect normal cells from the toxicity of chemotherapy drugs through p53 activation (cyclotherapy). An understanding of ActD's effect on p53 signaling is critical for the meaningful application of ActD in cyclotherapy-based combinations. This study evaluated the anti-tumor efficacy and mechanism of action of ActD in aerodigestive tract cancers. We found that ActD strongly inhibited the growth of a panel of aerodigestive tract cancer cell lines and induced efficient apoptosis, although the sensitivity varies among cell lines. The IC values of ActD spanned between 0.021 and 2.96 nM. Mechanistic studies revealed that ActD increased the expression of total and phosphorylated p53 (ser15) in a time- and dose-dependent manner. Moreover, ActD-induced apoptosis is dependent on p53 in cells expressing wild-type p53 and that ActD induced context-dependent differential expression of downstream targets p21 and PUMA without significant effects on p27. In the final analysis, this study revealed that p53-p21 is the predominant pathway activated by low-dose ActD, supporting further development of ActD in cyclotherapy.
Topics: Antibiotics, Antineoplastic; Apoptosis; Dactinomycin; Humans; Neoplasms; Tumor Suppressor Protein p53
PubMed: 35267106
DOI: 10.1007/s10495-022-01720-5 -
Inhibition of transcription by dactinomycin reveals a new characteristic of immunogenic cell stress.EMBO Molecular Medicine May 2020Chemotherapy still constitutes the standard of care for the treatment of most neoplastic diseases. Certain chemotherapeutics from the oncological armamentarium are able...
Chemotherapy still constitutes the standard of care for the treatment of most neoplastic diseases. Certain chemotherapeutics from the oncological armamentarium are able to trigger pre-mortem stress signals that lead to immunogenic cell death (ICD), thus inducing an antitumor immune response and mediating long-term tumor growth reduction. Here, we used an established model, built on artificial intelligence to identify, among a library of 50,000 compounds, anticancer agents that, based on their molecular descriptors, were predicted to induce ICD. This algorithm led us to the identification of dactinomycin (DACT, best known as actinomycin D), a highly potent cytotoxicant and ICD inducer that mediates immune-dependent anticancer effects in vivo. Since DACT is commonly used as an inhibitor of DNA to RNA transcription, we investigated whether other experimentally established or algorithm-selected, clinically employed ICD inducers would share this characteristic. As a common leitmotif, a panel of pharmacological ICD stimulators inhibited transcription and secondarily translation. These results establish the inhibition of RNA synthesis as an initial event for ICD induction.
Topics: Antineoplastic Agents; Artificial Intelligence; Dactinomycin; Humans; Immunogenic Cell Death; Neoplasms
PubMed: 32323922
DOI: 10.15252/emmm.201911622