Deregulacja mechanizmów epigenetycznych w nowotworach Marta Maleszewska*,

Maleszewska, Marta; Wojtaś, Bartosz; Kamińska, Bożena

doi:10.18388/pb.2018_125

Cited by 16 publications

(3 citation statements)

References 72 publications

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“…A remarkable complexity of chromatin organization, involving several levels of order of DNA coiling around the nucleosome cores and beyond the 30-nm "beads on a string" nucleosomal order, characterizes interphase chromatin and mitotic chromosomes (Mateo et al, 2019;Tan, Xing, Chang, Li, & Xie, 2018;Wang & Hayes, 2006;Bintu et al, 2018). Furthermore, considerable variability of chromatin organization characterizes different parts of genomes, various cell types, sequential developmental or differentiation stages of cells, and the respective cell cycle phases (Bannister & Kouzarides, 2011;Finn et al, 2019;Maleszewska, Wojtas, & Kamińska, 2018;Nir et al, 2018;Tolsma & Hansen, 2019). This variability comprises altered degrees of chromatin compaction (condensation) that represent both divergence in chromatin protein constituents and local epigenetic changes involving DNA and protein modifications such as phosphorylation, acetylation, methylation, among others.…”

Section: Of 14mentioning

confidence: 99%

Assessment of DNA Susceptibility to Denaturation as a Marker of Chromatin Structure

et al. 2019

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The susceptibility of DNA in situ to denaturation is modulated by its interactions with histone and nonhistone proteins, as well as with other chromatin components related to the maintenance of the 3D nuclear structure. Measurement of DNA proclivity to denature by cytometry provides insight into chromatin structure and thus can be used to recognize cells in different phases of the cell cycle, including mitosis, quiescence (G 0 ), and apoptosis, as well as to identify the effects of drugs that modify chromatin structure. Particularly useful is the method's ability to detect chromatin changes in sperm cells related to DNA fragmentation and infertility. This article presents a flow cytometric procedure for assessing DNA denaturation based on application of the metachromatic property of acridine orange (AO) to differentially stain singleversus double-stranded DNA. This approach circumvents limitations of biochemical methods of examining DNA denaturation, in particular the fact that the latter destroy higher orders of chromatin structure and that, being applied to bulk cell populations, they cannot detect heterogeneity of individual cells. Because the metachromatic properties of AO have also found application in other cytometric procedures, such as differential staining of RNA versus DNA and assessment of lysosomal proton pump including autophagy, to avert confusion between these approaches and the use of this dye in the DNA denaturation assay, these AO applications are briefly outlined in this unit as well. C 2019 by John Wiley & Sons, Inc. Basic Protocol: Differential staining of single-versus double-stranded DNA with acridine orange Keywords: acridine orange r apoptosis r autophagy r cell cycle r lysosomes r mitosis r sperm chromatin fertility assay How to cite this article: Darzynkiewicz, Z., Halicka, D. H., Zhao, H., & Li, J. (2019). Assessment of DNA susceptibility to denaturation as a marker of chromatin structure. BASIC PROTOCOL DNA DenaturationThe process of splitting double-stranded DNA (dsDNA) into single strands, which involves breaking hydrogen bonds between the bases in the DNA double-helical structure, is defined as DNA denaturation. DNA in solution undergoes denaturation when heated, and the temperature at which half remains in double-stranded conformation while the other half is single stranded is called the melting temperature (T m ). Because the G-C base pair, maintained by three hydrogen bonds, is stronger that the two-base pairing of AO has unique metachromatic properties: under the correct conditions (AO concentration, ionic strength, temperature), it can differentially stain ssDNA and dsDNA. Specifically, when AO intercalates between the base pairs of dsDNA, its fluorescence is maximally excited at 480 nm, its emission is at 530 nm, and its excitation lifetime is ß2 ns (Kapuscinski & Darzynkiewicz, 1984Kapuscinski, Darzynkiewicz, & Melamed, 1982. These values reflect singlet excitation of AO. The interaction of AO with ssDNA is more complex: it involves the insertion of the AO cation between adj...

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Section: Of 14mentioning

confidence: 99%

Assessment of DNA Susceptibility to Denaturation as a Marker of Chromatin Structure

et al. 2019

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“…It is generally accepted that, apart from genetic mutations in pro-oncogenes or tumor suppressor genes, epigenetic modifications may drive the oncogenic transformation [1,2]. The aberrant pattern of DNA methylation in neoplastic cells is often manifested as global hypomethylation accompanied by local hypermethylation of CpG islands.…”

Section: Introductionmentioning

confidence: 99%

The Contrasting Delayed Effects of Transient Exposure of Colorectal Cancer Cells to Decitabine or Azacitidine

Pawlak

Chybicka

Zioło

et al. 2022

Cancers

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(1) Background: Decitabine and azacitidine are cytosine analogues representing the class of drugs interfering with DNA methylation. Due to their molecular homology and similar clinical application, both drugs are often regarded as interchangeable. Despite their unique mechanism of action the studies designed for observation and comparison of the prolonged activity of these drugs are rare. (2) Methods: The short-time (20–72 h) and long-term (up to 20 days) anti-cancer activity of decitabine and azacitidine has been studied in colorectal cancer cells. We observe the impact on cell culture’s viability, clonogenicity, proliferation, and expression of CDKN1A, CCND1, MDM2, MYC, CDKN2A, GLB1 genes, and activity of SA-β-galactosidase. (3) Results: Decitabine has much stronger anti-clonogenic activity than azacitidine. We show that azacitidine, despite significant immediate toxicity, has negligible long-term effects. Contrary, decitabine, which does not exert initial toxicity, profoundly worsened the condition of the cells over time. On the 13th day after treatment, the viability of cells was decreased and proliferation inhibited. These functional changes were accompanied by up-regulation of expression CDKN1A, CCND1, and CDKN2A genes and increased activation of SA-β-galactosidase, indicating cellular senescence. (4) Conclusions: Our head-to-head comparison revealed profound differences in the activities of decitabine and azacitidine important in their anti-cancer potential and clinical application. The effects of decitabine need relatively long time to develop. This property is crucial for proper design of studies and therapy concerning decitabine and undermines opinion about the similar therapeutic mechanism and interchangeability of these drugs.

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“…Epigenetic modifiers are widely recognized as targets for therapeutic intervention, owing to their critical roles in regulating gene expression and chromatin integrity in addition to their dysregulation in a range of human pathologies 3. In particular, the nuclear receptor-binding SET domain NSD2 family of histone lysine methyltransferase enzymes, namely NSD1, NSD2/WHSC1/MMSET, and NSD3/WHSC1L1, which are involved in developmental and tumoral pathologies,4 have all been implicated as cancer therapeutic targets 5.…”

Section: Introductionmentioning

confidence: 99%

<p>WHSC1 acts as a prognostic indicator and functions as an oncogene in cervical cancer</p>

Luo

Duan

et al. 2019

OTT

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Purpose: Wolf–Hirschhorn syndrome candidate 1 (WHSC1) is an epigenetic modifier, considered to play a driving role in oncogenesis. However, very little is known about the roles of WHSC1 and its prognostic impacts in cervical cancer. This study aimed to investigate the role of WHSC1 in the prognosis of cervical cancer and explore the effect of WHSC1 on proliferation, migration, and invasion of cervical cancer cells and angiogenesis in human umbilical vein endothelial cells (HUVECs). Methods: We evaluated the expression levels of WHSC1 in cervical cancer samples and relevant cell lines by immunohistochemistry, real-time quantitative PCR, and Western blot. In vitro, Cell Counting Kit-8 and transwell assays were used to investigate the viability and migration of C33A cells, and a tube formation assay was used to study the effect of WHSC1 on angiogenesis in HUVECs. Results: WHSC1 was overexpressed in cervical cancer tissues and cells, and correlated with the FIGO stage and differentiation. WHSC1 knockdown inhibited proliferation, suppressed migration and invasion in endothelial nitric oxide synthase (eNOS)-overexpressing C33A cells, and inhibited angiogenesis in HUVECs. Conclusion: WHSC1 may be a poor prognostic indicator of cervical cancer and a potential novel therapeutic target for the same. WHSC1 may participate in the regulation of cervical cancer progression through the eNOS signaling pathway.

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Deregulacja mechanizmów epigenetycznych w nowotworach Marta Maleszewska*,

Cited by 16 publications

References 72 publications

Assessment of DNA Susceptibility to Denaturation as a Marker of Chromatin Structure

Assessment of DNA Susceptibility to Denaturation as a Marker of Chromatin Structure

The Contrasting Delayed Effects of Transient Exposure of Colorectal Cancer Cells to Decitabine or Azacitidine

<p>WHSC1 acts as a prognostic indicator and functions as an oncogene in cervical cancer</p>

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