Transcription of carbonyl reductase 1 is regulated by DNA topoisomerase II beta

Khazeem, Mushtaq M; Cowell, Ian G.; Harkin, Lauren F.; Casement, John; Austin, Caroline A.

doi:10.1002/1873-3468.13904

Cited by 3 publications

(5 citation statements)

References 43 publications

(95 reference statements)

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“…Cells were grown in 96-well plates, and RNA samples used for RT-qPCR were prepared directly from fresh cell lysates using SingleShot Cell Lysis Kit (Bio-Rad) according to manufacturer's instructions as previously described [49].…”

Section: Rt-pcrmentioning

confidence: 99%

“…VCA-1003) according to the manufacturer's instructions. Cells were selected and sorted based on GFP expression as described previously [49]. GFP-positive cells were incubated at 37 °C for 2-3 weeks until colonies formed.…”

Section: Rt-pcrmentioning

confidence: 99%

“…Four biological replicate RNA samples were prepared for each of the four conditions. RNA extraction, quantification, and quality checking were as previously described [49]. Library preparation employed TruSeq-stranded mRNA kit (Illumina) which selects only poly-A containing mRNA molecules using poly-T oligo attached magnetic beads using the High Sample (HS) Protocol of (TruSeq® Stranded mRNA Sample Preparation Guide, Illumina) was followed.…”

Section: Rna-seqmentioning

confidence: 99%

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TOP2B Is Required to Maintain the Adrenergic Neural Phenotype and for ATRA-Induced Differentiation of SH-SY5Y Neuroblastoma Cells

et al. 2022

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The neuroblastoma cell line SH-SY5Y is widely used to study retinoic acid (RA)-induced gene expression and differentiation and as a tool to study neurodegenerative disorders. SH-SY5Y cells predominantly exhibit adrenergic neuronal properties, but they can also exist in an epigenetically interconvertible alternative state with more mesenchymal characteristics; as a result, these cells can be used to study gene regulation circuitry controlling neuroblastoma phenotype. Using a combination of pharmacological inhibition and targeted gene inactivation, we have probed the requirement for DNA topoisomerase IIB (TOP2B) in RA-induced gene expression and differentiation and in the balance between adrenergic neuronal versus mesenchymal transcription programmes. We found that expression of many, but not all genes that are rapidly induced by ATRA in SH-SY5Y cells was significantly reduced in the TOP2B null cells; these genes include BCL2, CYP26A1, CRABP2, and NTRK2. Comparing gene expression profiles in wild-type versus TOP2B null cells, we found that long genes and genes expressed at a high level in WT SH-SY5Y cells were disproportionately dependent on TOP2B. Notably, TOP2B null SH-SY5Y cells upregulated mesenchymal markers vimentin (VIM) and fibronectin (FN1) and components of the NOTCH signalling pathway. Enrichment analysis and comparison with the transcription profiles of other neuroblastoma-derived cell lines supported the conclusion that TOP2B is required to fully maintain the adrenergic neural-like transcriptional signature of SH-SY5Y cells and to suppress the alternative mesenchymal epithelial-like epigenetic state.

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Section: Rt-pcrmentioning

confidence: 99%

Section: Rt-pcrmentioning

confidence: 99%

Section: Rna-seqmentioning

confidence: 99%

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TOP2B Is Required to Maintain the Adrenergic Neural Phenotype and for ATRA-Induced Differentiation of SH-SY5Y Neuroblastoma Cells

et al. 2022

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“…Combination indexes were determined using CalcuSyn. HL-60 homozygous mutants (TOP2B −/− ) and the corresponding wild-type controls (TOP2B +/+ ) prepared using CRISPR-Cas9 technology as described previously ( Khazeem et al , 2020 , 2022 ) and briefly in Supplementary Material (Section 1.2) were incubated in IMDM media (Gibco) supplemented with 10% FBS, and were processed by the same procedure for the viability assay.…”

Section: Methodsmentioning

confidence: 99%

Exploring the effects of topoisomerase II inhibitor XK469 on anthracycline cardiotoxicity and DNA damage

Keresteš,

Kubeš,

Applová

et al. 2024

Toxicological Sciences

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Anthracyclines, such as doxorubicin (adriamycin), daunorubicin, or epirubicin, rank among the most effective agents in classical anticancer chemotherapy. However, cardiotoxicity remains the main limitation of their clinical use. Topoisomerase IIβ has recently been identified as a plausible target of anthracyclines in cardiomyocytes. We examined the putative topoisomerase IIβ selective agent XK469 as a potential cardioprotective and designed several new analogs. In our experiments, XK469 inhibited both topoisomerase isoforms (α and β) and did not induce topoisomerase II covalent complexes in isolated cardiomyocytes and HL-60, but induced proteasomal degradation of topoisomerase II in these cell types. The cardioprotective potential of XK469 was studied on rat neonatal cardiomyocytes, where dexrazoxane (ICRF-187), the only clinically approved cardioprotective, was effective. Initially, XK469 prevented daunorubicin-induced toxicity and p53 phosphorylation in cardiomyocytes. However, it only partially prevented the phosphorylation of H2AX and did not affect DNA damage measured by Comet Assay. It also did not compromise the daunorubicin antiproliferative effect in HL-60 leukemic cells. When administered to rabbits to evaluate its cardioprotective potential in vivo, XK469 failed to prevent the daunorubicin-induced cardiac toxicity in either acute or chronic settings. In the following in vitro analysis, we found that prolonged and continuous exposure of rat neonatal cardiomyocytes to XK469 led to significant toxicity. In conclusion, this study provides important evidence on the effects of XK469 and its combination with daunorubicin in clinically relevant doses in cardiomyocytes. Despite its promising characteristics, long-term treatments and in vivo experiments have not confirmed its cardioprotective potential.

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“…VCA-1003, for SH-SY5Y and K562 cells) according to the manufacturer's instructions. Cells were selected and sorted based on GFP expression as reported previously ( 48 ) as a single cell per well into 96-well plates and incubated at 37°C for 2–3 weeks until colonies formed. Colonies were expanded, and screening for TOP2B null clones was performed using genotyping and immunofluorescence as in ( 48 ).…”

Section: Methodsmentioning

confidence: 99%

Two type I topoisomerases maintain DNA topology in human mitochondria

Menger

Chapman

Díaz-Maldonado

et al. 2022

Nucleic Acids Research

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Genetic processes require the activity of multiple topoisomerases, essential enzymes that remove topological tension and intermolecular linkages in DNA. We have investigated the subcellular localisation and activity of the six human topoisomerases with a view to understanding the topological maintenance of human mitochondrial DNA. Our results indicate that mitochondria contain two topoisomerases, TOP1MT and TOP3A. Using molecular, genomic and biochemical methods we find that both proteins contribute to mtDNA replication, in addition to the decatenation role of TOP3A, and that TOP1MT is stimulated by mtSSB. Loss of TOP3A or TOP1MT also dysregulates mitochondrial gene expression, and both proteins promote transcription elongation in vitro. We find no evidence for TOP2 localisation to mitochondria, and TOP2B knockout does not affect mtDNA maintenance or expression. Our results suggest a division of labour between TOP3A and TOP1MT in mtDNA topology control that is required for the proper maintenance and expression of human mtDNA.

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Transcription of carbonyl reductase 1 is regulated by DNA topoisomerase II beta

Cited by 3 publications

References 43 publications

TOP2B Is Required to Maintain the Adrenergic Neural Phenotype and for ATRA-Induced Differentiation of SH-SY5Y Neuroblastoma Cells

TOP2B Is Required to Maintain the Adrenergic Neural Phenotype and for ATRA-Induced Differentiation of SH-SY5Y Neuroblastoma Cells

Exploring the effects of topoisomerase II inhibitor XK469 on anthracycline cardiotoxicity and DNA damage

Two type I topoisomerases maintain DNA topology in human mitochondria

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