Evidence map›Paper›PMID 38892135›Full record

ArticleInternational journal of molecular sciences2024

Cellular Distribution and Ultrastructural Changes in HaCaT Cells, Induced by Podophyllotoxin and Its Novel Fluorescent Derivative, Supported by the Molecular Docking Studies.

Piotr Strus, Karol Sadowski, Julia Kostro, Andrzej Antoni Szczepankiewicz, Hanna Nieznańska, Magdalena Niedzielska, Andrei Zlobin, Pramukti Nawar Ra'idah, Zuzanna Molęda, Joanna Szawkało and 4 more

Abstract read
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In one paragraph

Article in International journal of molecular sciences, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed, 1 pooled it
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 1 synthesis or guideline pooled it.

  1. Pooled it
  2. Article
  3. Article
  4. Article
  5. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

14 authors.

Piotr StrusDepartment of Histology and Embryology, Medical University of Warsaw, Chałubińskiego 5, 02-004 Warsaw, Poland.ORCID 0000-0003-2246-0481
Karol SadowskiStudents Scientific Group HESA, Department of Histology and Embryology, Medical University of Warsaw, Chałubinskiego 5, 02-004 Warsaw, Poland.
Julia KostroStudents Scientific Group HESA, Department of Histology and Embryology, Medical University of Warsaw, Chałubinskiego 5, 02-004 Warsaw, Poland.
Andrzej Antoni SzczepankiewiczLaboratory of Electron Microscopy, Nencki Institute of Warsaw, Pasteura 3, 02-093 Warsaw, Poland.ORCID 0000-0002-1502-3147
Hanna NieznańskaLaboratory of Electron Microscopy, Nencki Institute of Warsaw, Pasteura 3, 02-093 Warsaw, Poland.
Magdalena NiedzielskaLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.ORCID 0000-0001-9618-7970
Andrei ZlobinLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.
Pramukti Nawar Ra'idahLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.
Zuzanna MolędaLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.ORCID 0000-0002-0868-2216
Joanna SzawkałoLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.
Zbigniew CzarnockiLaboratory of Natural Products Chemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.ORCID 0000-0002-6581-9282
Cezary WójcikAmgen Inc., Thousand Oaks, CA 91320, USA.ORCID 0000-0003-2509-7251
Łukasz SzeleszczukDepartment of Organic and Physical Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-093 Warsaw, Poland.ORCID 0000-0002-5302-3797
Izabela Młynarczuk-BiałyDepartment of Histology and Embryology, Medical University of Warsaw, Chałubińskiego 5, 02-004 Warsaw, Poland.ORCID 0000-0002-5192-6415

Funding

Medical University of Warsaw 1M15/1/MB/N/22-23 to PSPolish Ministry for Education and Science MEiN/2022/DIR/3155
6 · The paper itself

Abstract

Podophyllotoxin (PPT) is an active pharmaceutical ingredient (API) with established antitumor potential. However, due to its systemic toxicity, its use is restricted to topical treatment of anogenital warts. Less toxic PPT derivatives (e.g., etoposide and teniposide) are used intravenously as anticancer agents. PPT has been exploited as a scaffold of new potential therapeutic agents; however, fewer studies have been conducted on the parent molecule than on its derivatives. We have undertaken a study of ultrastructural changes induced by PPT on HaCaT keratinocytes. We have also tracked the intracellular localization of PPT using its fluorescent derivative (PPT-FL). Moreover, we performed molecular docking of both PPT and PPT-FL to compare their affinity to various binding sites of tubulin. Using the Presto blue viability assay, we established working concentrations of PPT in HaCaT cells. Subsequently, we have used selected concentrations to determine PPT effects at the ultrastructural level. Dynamics of PPT distribution by confocal microscopy was performed using PPT-FL. Molecular docking calculations were conducted using Glide. PPT induces a time-dependent cytotoxic effect on HaCaT cells. Within 24 h, we observed the elongation of cytoplasmic processes, formation of cytoplasmic vacuoles, progressive ER stress, and shortening of the mitochondrial long axis. After 48 h, we noticed disintegration of the cell membrane, progressive vacuolization, apoptotic/necrotic vesicles, and a change in the cell nucleus's appearance. PPT-FL was detected within HaCaT cells after ~10 min of incubation and remained within cells in the following measurements. Molecular docking confirmed the formation of a stable complex between tubulin and both PPT and PPT-FL. However, it was formed at different binding sites. PPT is highly toxic to normal human keratinocytes, even at low concentrations. It promptly enters the cells, probably via endocytosis. At lower concentrations, PPT causes disruptions in both ER and mitochondria, while at higher concentrations, it leads to massive vacuolization with subsequent cell death. The novel derivative of PPT, PPT-FL, forms a stable complex with tubulin, and therefore, it is a useful tracker of intracellular PPT binding and trafficking.

Indexed as

HaCaT CellsKeratinocytesMolecular Docking SimulationPodophyllotoxinTubulinBinding SitesCell SurvivalEndoplasmic Reticulum StressFluorescent DyesHumansMitochondriaFluorescent DyesPodophyllotoxinTubulincell deathHaCaT ultrastructurehuman keratinocytespodophyllotoxinpodophyllotoxin docking to tubulinpodophyllotoxin with attached fluorescein

Identifiers

PMID38892135
PMCPMC11172492

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.