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Environmental Toxicology Mar 2024The treatment of clear-cell renal cell carcinoma (ccRCC) remains challenge. Chemokines laid impact on the proliferation and metastasis of cancer cells. The objective was...
BACKGROUND
The treatment of clear-cell renal cell carcinoma (ccRCC) remains challenge. Chemokines laid impact on the proliferation and metastasis of cancer cells. The objective was to identify the chemokine-related genes and construct a prognostic model for ccRCC.
METHODS
Bulk transcriptomic data (n = 531), single-cell RNA sequencing (scRNA-seq) dataset GSE159115, and other validation cohorts were acquired from the Cancer Genome Atlas Program (TCGA) and GEO databases. All clustering analysis was conducted by Seurat R package. Gene set enrichment analysis (GSEA), immune infiltration analysis, single nucleotide variations (SNV) analysis, and predictive response analysis of immunotherapy/chemotherapy were conducted. 786-O and A498 cell lines were cultured and applied into CCK-8, Western blot, and RT-qPCR kits.
RESULTS
Univariate Cox analysis was used to screen out chemokine-related genes related to survival. ZIC2, SMIM24, COL7A1, IGF2BP3, ITPKA, ADAMTS14, CYP3A7, and AURKB were identified and applied for the construction of the prognostic model. High-risk group had a poorer prognosis than the low-risk group in each dataset. Memory CD8+ T cells, macrophages, and memory B cells were higher in the high-risk group, while the content of basophils was higher in the low-risk group. Bortezomib_1191, Dactinomycin_1911, Docetaxel_1007, and Daporinad_1248 were more sensitive to high-risk groups than low-risk groups. Moreover, we found that IGF2BP3 significantly elevated in both 786-O and A498 cell lines resistance to sunitinib. Knockdown of IGF2BP3 markedly reduced ccRCC cell migration and viability.
CONCLUSION
Our study has yielded a novel prognostic model of chemokine-related genes based on comprehensive transcriptional atlas of ccRCC patients, shedding light on the significant impact of the tumor microenvironment on biology and immunotherapy response of ccRCC. We identified IGF2BP3 as a pivotal regulator in regulating ccRCC resistance to sunitinib.
PubMed: 38488671
DOI: 10.1002/tox.24190 -
JCEM Case Reports Mar 2024Thyroid storm due to gestational trophoblastic disease (GTD) with metastatic choriocarcinoma is a rare but potentially life-threatening endocrine emergency. We report on...
Thyroid storm due to gestational trophoblastic disease (GTD) with metastatic choriocarcinoma is a rare but potentially life-threatening endocrine emergency. We report on a woman with molar pregnancy and metastatic choriocarcinoma who presented with thyroid storm (Burch-Wartofsky point scale of 45) a few weeks after the evacuation of GTD. She was initially managed with intravenous hydrocortisone, oral propylthiouracil (PTU), and esmolol infusion. After stabilization in the intensive care unit, 10 cycles of chemotherapy with etoposide, methotrexate, leucovorin, dactinomycin, and cyclophosphamide (EMA-CO) were initiated for stage 4 choriocarcinoma with brain and lung metastases. She underwent a hysterectomy soon after completing chemotherapy and received an additional 3 cycles of chemotherapy after the hysterectomy. As human chorionic gonadotropin (hCG) levels normalized, thyroid function reverted to normal as well. At the last follow-up, the patient was asymptomatic, euthyroid (without antithyroid medication), had a normal hCG titer of 1.7 mIU/mL (normal nonpregnant reference is < 5 mIU/mL), and the lung and brain lesions had resolved entirely. Management of thyroid storm in the presence of untreated metastatic choriocarcinoma requires a high index of suspicion and a multidisciplinary team approach to prevent complications and improve survival.
PubMed: 38476634
DOI: 10.1210/jcemcr/luae019 -
Chemico-biological Interactions Apr 2024Transcriptomic analyses have revealed hundreds of p53-regulated genes; however, these studies used a limited number of cell lines and p53-activating agents. Therefore,...
Transcriptomic analyses have revealed hundreds of p53-regulated genes; however, these studies used a limited number of cell lines and p53-activating agents. Therefore, we searched for candidate p53-target genes by employing stress factors and cell lines never before used in a high-throughput search for p53-regulated genes. We performed RNA-Seq on A549 cells exposed to camptothecin, actinomycin D, nutlin-3a, as well as a combination of actinomycin D and nutlin-3a (A + N). The latter two substances synergise upon the activation of selected p53-target genes. A similar analysis was performed on other cell lines (U-2 OS, NCI-H460, A375) exposed to A + N. To identify proteins in cell lysates or those secreted into a medium of A549 cells in control conditions or treated with A + N, we employed mass spectrometry. The expression of selected genes strongly upregulated by A + N or camptothecin was examined by RT-PCR in p53-deficient cells and their controls. We found that p53 participates in the upregulation of: ACP5, APOL3, CDH3, CIBAR2, CRABP2, CTHRC1, CTSH, FAM13C, FBXO2, FRMD8, FRZB, GAST, ICOSLG, KANK3, KCNK6, KLRG2, MAFB, MR1, NDRG4, PTAFR, RETSAT, TMEM52, TNFRSF14, TRANK1, TYSND1, WFDC2, WFDC5, WNT4 genes. Twelve of these proteins were detected in the secretome and/or proteome of treated cells. Our data generated new hypotheses concerning the functioning of p53. Many genes activated by A + N or camptothecin are also activated by interferons, indicating a noticeable overlap between transcriptional programs of p53 and these antiviral cytokines. Moreover, several identified genes code for antagonists of WNT/β-catenin signalling pathways, which suggests new connections between these two cancer-related signalling systems. One of these antagonists is DRAXIN. Previously, we found that its gene is activated by p53. In this study, using mass spectrometry and Western blotting, we detected expression of DRAXIN in a medium of A549 cells exposed to A + N. Thus, this protein functions not only in the development of the nervous system, but it may also have a new cancer-related function.
Topics: Dactinomycin; Tumor Suppressor Protein p53; Cell Line, Tumor; Proteomics; Camptothecin; Gene Expression Profiling; Apoptosis; Neoplasms; Imidazoles; Piperazines
PubMed: 38460933
DOI: 10.1016/j.cbi.2024.110946 -
Investigative Ophthalmology & Visual... Feb 2024N4-acetylcytidine (ac4C) is a post-transcriptional RNA modification catalyzed by N-acetyltransferase 10 (NAT10), a critical factor known to influence mRNA stability....
PURPOSE
N4-acetylcytidine (ac4C) is a post-transcriptional RNA modification catalyzed by N-acetyltransferase 10 (NAT10), a critical factor known to influence mRNA stability. However, the role of ac4C in visual development remains unexplored.
METHODS
Analysis of public datasets and immunohistochemical staining were conducted to assess the expression pattern of nat10 in zebrafish. We used CRISPR/Cas9 and RNAi technologies to knockout (KO) and knockdown (KD) nat10, the zebrafish ortholog of human NAT10, and evaluated its effects on early development. To assess the impact of nat10 knockdown on visual function, we performed comprehensive histological evaluations and behavioral analyses. Transcriptome profiling and real-time (RT)-PCR were utilized to detect alterations in gene expression resulting from the nat10 knockdown. Dot-blot and RNA immunoprecipitation (RIP)-PCR analyses were conducted to verify changes in ac4C levels in both total RNA and opsin mRNA specifically. Additionally, we used the actinomycin D assay to examine the stability of opsin mRNA following the nat10 KD.
RESULTS
Our study found that the zebrafish NAT10 protein shares similar structural properties with its human counterpart. We observed that the nat10 gene was prominently expressed in the visual system during early zebrafish development. A deficiency of nat10 in zebrafish embryos resulted in increased mortality and developmental abnormalities. Behavioral and histological assessments indicated significant vision impairment in nat10 KD zebrafish. Transcriptomic analysis and RT-PCR identified substantial downregulation of retinal transcripts related to phototransduction, light response, photoreceptors, and visual perception in the nat10 KD group. Dot-blot and RIP-PCR analyses confirmed a pronounced reduction in ac4C levels in both total RNA and specifically in opsin messenger RNA (mRNA). Additionally, by evaluating mRNA decay in zebrafish treated with actinomycin D, we observed a significant decrease in the stability of opsin mRNA in the nat10 KD group.
CONCLUSIONS
The ac4C-mediated mRNA modification plays an essential role in maintaining visual development and retinal function. The loss of NAT10-mediated ac4C modification results in significant disruptions to these processes, underlining the importance of this RNA modification in ocular development.
Topics: Humans; Animals; Acetyltransferases; Zebrafish; Dactinomycin; Opsins; Rod Opsins; RNA; RNA, Messenger
PubMed: 38381411
DOI: 10.1167/iovs.65.2.31 -
Journal of Agricultural and Food... Mar 2024The present study investigated the antibacterial mechanism, control efficiency, and nontarget toxicity of actinomycin X (Act-X) against subsp. citri (Xcc) for the first...
The present study investigated the antibacterial mechanism, control efficiency, and nontarget toxicity of actinomycin X (Act-X) against subsp. citri (Xcc) for the first time. Act-X almost completely inhibited the proliferation of Xcc in the growth curve assay at a concentration of 0.25 MIC (minimum inhibitory concentration, MIC = 31.25 μg/mL). This inhibitory effect was achieved by increasing the production of reactive oxygen species (ROS), blocking the formation of biofilms, obstructing the synthesis of intracellular proteins, and decreasing the enzymatic activities of malate dehydrogenase (MDH) and succinate dehydrogenase (SDH) of Xcc. Molecular docking and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis results indicated that Act-X steadily bonded to the RNA polymerase, ribosome, malate dehydrogenase, and succinate dehydrogenase to inhibit their activities, thus drastically reducing the expression levels of related genes. Act-X showed far more effectiveness than the commercially available pesticide Cu(OH)Cl in the prevention and therapy of citrus canker disease. Furthermore, the nontarget toxicity evaluation demonstrated that Act-X was not phytotoxic to citrus trees and exhibited minimal toxicity to earthworms in both contact and soil toxic assays. This study suggests that Act-X has the potential as an effective and environmentally friendly antibacterial agent.
Topics: Malate Dehydrogenase; Succinate Dehydrogenase; Molecular Docking Simulation; Anti-Bacterial Agents; Citrus; Xanthomonas; Plant Diseases; Dactinomycin
PubMed: 38377546
DOI: 10.1021/acs.jafc.3c08600 -
Case Reports in Women's Health Mar 2024This case report outlines the clinical course of a young woman who presented as haemodynamically unstable due to uterine perforation. She had undergone suction dilation...
This case report outlines the clinical course of a young woman who presented as haemodynamically unstable due to uterine perforation. She had undergone suction dilation and curettage three weeks prior and received a diagnosis of complete molar pregnancy. During her most recent acute presentation, an emergency laparotomy revealed a full-thickness fundal uterine rupture in a region of newly identified arteriovenous malformation. Haemostasis was achieved with the primary repair of the perforation. She was subsequently diagnosed with gestational trophoblastic neoplasm (GTN), a condition characterised by abnormal proliferation of trophoblastic tissue. She received three courses of methotrexate followed by a two-month course of dactinomycin. At one-year surveillance, she had made a complete recovery.
PubMed: 38312237
DOI: 10.1016/j.crwh.2023.e00579 -
British Journal of Haematology Apr 2024We have conducted a retrospective, single-centre analysis of 20 patients with relapsed or refractory FLT3-mutated acute myeloid leukaemia (FLT3m AML) who received a...
We have conducted a retrospective, single-centre analysis of 20 patients with relapsed or refractory FLT3-mutated acute myeloid leukaemia (FLT3m AML) who received a salvage quadruplet regimen consisting of gilteritinib, venetoclax, low-dose cytarabine and actinomycin D (G-ACTIVE). G-ACTIVE resulted in a 95% (19/20) overall response rate and 75% (15/20) complete remission and complete remission with an incomplete platelet recovery (CR + CRp) rate. Out of 13 transplant-eligible patients, 11 (86%) proceeded to an allogeneic stem cell transplantation. The median overall survival and relapse-free survival after G-ACTIVE were 32 and 12.9 months respectively. The Day 60 mortality rate was 15%.
Topics: Humans; Cytarabine; Dactinomycin; Retrospective Studies; Neoplasm Recurrence, Local; Leukemia, Myeloid, Acute; fms-Like Tyrosine Kinase 3; Aniline Compounds; Pyrazines; Sulfonamides; Bridged Bicyclo Compounds, Heterocyclic
PubMed: 38291741
DOI: 10.1111/bjh.19318 -
Life Sciences Mar 2024Although the functions of progesterone in the myometrium are well-established, the nongenomic effects of progesterone in pregnant myometrial contractions are still...
AIMS
Although the functions of progesterone in the myometrium are well-established, the nongenomic effects of progesterone in pregnant myometrial contractions are still unclear. Therefore, this study aimed to investigate changes in the nongenomic effects of progesterone during pregnancy.
MAIN METHODS
Myometrial strips were obtained from non-pregnant, pregnant, and postpartum rats, and the nongenomic effects of progesterone in the myometrium during pregnancy were examined. Additionally, the influence of actinomycin D and cycloheximide and the effects of Org OD-02-0 (a specific membrane progesterone receptor (mPR) agonist) in the myometrium were investigated. Moreover, DNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to identify genes involved in progesterone-induced effects in the myometrium.
KEY FINDINGS
Progesterone did not cause rhythmic contractions in non-pregnant myometrium but induced rhythmic contractions in pregnant myometrium, with the effects peaking at 20 d + 8 h of pregnancy. However, myometrial contractions decreased after delivery and were restored to non-pregnant levels at 7 d postpartum. Additionally, progesterone stably inhibited high KCl-induced myometrial contractions during pregnancy. Moreover, the nongenomic effects of progesterone were unaffected by actinomycin D or cycloheximide, and Org OD-02-0 effectively mimicked these effects. DNA microarray analysis and qRT-PCR revealed a significant increase in mPRβ gene expression during pregnancy. However, mPRα, mPRγ, mPRδ, and mPRε expression levels remained unchanged.
SIGNIFICANCE
The stimulatory nongenomic effect of progesterone, which was inducible and mPRβ-dependent during pregnancy, may be involved in parturition. The inhibitory effect, which was constitutive and depended on other mPRs, may be involved in pregnancy maintenance.
Topics: Pregnancy; Female; Rats; Animals; Progesterone; Myometrium; Cycloheximide; Dactinomycin; Receptors, Progesterone; Progestins; Uterine Contraction
PubMed: 38262574
DOI: 10.1016/j.lfs.2024.122454 -
Pesticide Biochemistry and Physiology Jan 2024Citrus blue and green molds caused by Penicillium digitatum, P. italicum, and P. polonicum, are the major postharvest diseases of citrus fruit. In the present study,...
Citrus blue and green molds caused by Penicillium digitatum, P. italicum, and P. polonicum, are the major postharvest diseases of citrus fruit. In the present study, Actinomycin X (Act-X), a naturally occurring antibiotic produced by Streptomyces species, was found to show excellent antifungal effect against these three pathogens with a minimum inhibitory concentration (MIC) value of 62.5 μg/mL for them all, which was better than the positive control thiophanate-methyl. Act-X significantly reduced the percentage of spore germination, and highly inhibited the mycelial growth of P. italicum, P. digitatum, and P. polonicum with EC values being 34.34, 13.76, and 37.48 μg/mL, respectively. In addition, Act-X greatly decreased the intracellular protein content while increasing the reactive oxygen species (ROS) level and superoxide anion (O) content in the mycelia of pathogens. In vivo test indicated that Act-X strongly inhibited the infection of navel oranges by these three Penicillium species, with an inhibition percentage of >50% for them all at the concentration of 10 MIC. Transcriptome analysis suggested that Act-X might highly influence the ribosomal functions of P. polonicum, which was supported as well by the molecular docking analysis of Act-X with some key functional proteins and RNAs of the ribosome. Furthermore, Act-X significantly reduced the decay percentage and improved the firmness, color, and sugar-acid ratio of navel oranges spray-inoculated with P. polonicum during the postharvest storage at 4 °C for 60 d.
Topics: Antifungal Agents; Citrus; Molecular Docking Simulation; Fungi; Fruit; Dactinomycin
PubMed: 38225074
DOI: 10.1016/j.pestbp.2023.105718 -
SLAS Discovery : Advancing Life... Mar 2024Combination therapies have improved outcomes for patients with acute myeloid leukemia (AML). However, these patients still have poor overall survival. Although many...
Combination therapies have improved outcomes for patients with acute myeloid leukemia (AML). However, these patients still have poor overall survival. Although many combination therapies are identified with high-throughput screening (HTS), these approaches are constrained to disease models that can be grown in large volumes (e.g., immortalized cell lines), which have limited translational utility. To identify more effective and personalized treatments, we need better strategies for screening and exploring potential combination therapies. Our objective was to develop an HTS platform for identifying effective combination therapies with highly translatable ex vivo disease models that use size-limited, primary samples from patients with leukemia (AML and myelodysplastic syndrome). We developed a system, ComboFlow, that comprises three main components: MiniFlow, ComboPooler, and AutoGater. MiniFlow conducts ex vivo drug screening with a miniaturized flow-cytometry assay that uses minimal amounts of patient sample to maximize throughput. ComboPooler incorporates computational methods to design efficient screens of pooled drug combinations. AutoGater is an automated gating classifier for flow cytometry that uses machine learning to rapidly analyze the large datasets generated by the assay. We used ComboFlow to efficiently screen more than 3000 drug combinations across 20 patient samples using only 6 million cells per patient sample. In this screen, ComboFlow identified the known synergistic combination of bortezomib and panobinostat. ComboFlow also identified a novel drug combination, dactinomycin and fludarabine, that synergistically killed leukemic cells in 35 % of AML samples. This combination also had limited effects in normal, hematopoietic progenitors. In conclusion, ComboFlow enables exploration of massive landscapes of drug combinations that were previously inaccessible in ex vivo models. We envision that ComboFlow can be used to discover more effective and personalized combination therapies for cancers amenable to ex vivo models.
Topics: Humans; Drug Synergism; Drug Combinations; Leukemia, Myeloid, Acute; Panobinostat; Hematologic Neoplasms
PubMed: 38101570
DOI: 10.1016/j.slasd.2023.12.001