Morphological analysis and cytotoxicity of acrylamide on SPC212 human mesothelioma cells: Do low doses induce proliferation, while high doses cause toxicity?

Our plain-English summary of Morphological analysis and cytotoxicity of acrylamide on SPC212 human mesothelioma cells: Do low doses induce proliferation, while high doses cause toxicity?, by S. Kacar, O. Tomsuk (Journal of Cellular and Molecular Medicine, 2024).

The study

• Authors: S. Kacar, O. Tomsuk
• Journal: Journal of Cellular and Molecular Medicine
• Published: 2024;28(21):e70190
• Cited by: 1 article, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Acrylamide is broadly utilized in numerous areas with different purposes including being an additive, flocculating, sealing, dry strength improver and polymerizing agent, and so forth. Furthermore, it forms in certain food products at high temperatures. It poses serious hazard since its readily water-soluble and very reactive nature. Besides in vivo studies, several in vitro studies with various cell lines are carried out to evaluate its toxicity. However, of these cell line studies, there are no mesothelium or mesothelioma cell lines. To fill this lacuna, we aimed at examining various dose range of acrylamide on SPC212 human mesothelioma cell line. First, we executed MTT and neutral red cytotoxicity tests and ascertained IC50 dose. Next, we performed inverted, light (haematoxylin-eosin and May Grünwald), fluorescent (DAPI) and confocal microscope (AO/EB) analyses as well as immunohistochemistry for Bax, Bcl-2 and PCNA proteins. As a result, we found IC50 of acrylamide at 2.65 mM. Starting from 3.13 mM of acrylamide dose, a deep decrease in cell proliferation was observed. Particularly in MTT assay, a proliferative action of acrylamide was detected at 0.39 and 0.78 mM, supported with inverted microscope images. In light microscope analysis, several cellular degenerations, including condensed and kidney-shaped nucleus were evident. In AO/EB staining, cells with apoptotic characteristics augmented dose-dependently, being upheld by a parallel uptick in Bax and a dimunition in Bcl-2 staining. Besides, PCNA decreased at IC50 dose of acrylamide. This is the acrylamide-associated first study conducted on SPC212 mesothelioma cells encompassing advanced morphological analysis. We believe this study to be an incentive for future studies.

© 2024 The Author(s). Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.

In plain English

This laboratory experiment used SPC212, a human mesothelioma cell line from the pleural cavity of a 47-year-old woman. The authors exposed the cells to the chemical acrylamide for 24 hours at 0.39 to 25 mM. Asbestos was not studied. In the MTT viability test, the two lowest doses (0.39 and 0.78 mM) gave higher viability than untreated cells: 122.1% and 119.8%. Viability fell to 31.2% at 3.13 mM, 36.8% at 6.25 mM, 27.9% at 12.50 mM and 18.0% at 25 mM. In the neutral red test, the two lowest doses showed no significant change; viability fell to 73.4% at 1.56 mM and 13.5% at 3.13 mM. Based on the MTT test, the dose halving viability (IC50) was 2.65 mM. Under the microscope, treated cells showed shrinkage, condensed nuclei and other changes the authors linked to apoptosis (programmed cell death). At 2.00 mM, Bax staining increased while Bcl-2 and PCNA staining decreased. The authors concluded that in this cell line acrylamide was proliferative at low doses and inhibiting at higher doses. They noted they did not run further proliferation tests.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: in vitro laboratory experiment on a human mesothelioma cell line (cytotoxicity tests, oxidative stress measurements, microscopy and immunocytochemistry)
• Who was studied: SPC212 human mesothelioma cell line, described as obtained from the pleural cavity of a 47-year-old woman; no human subjects or animals were studied
• Exposure: Acrylamide (not asbestos) applied to cells for 24 hours at doses from 0.39 to 25 mM. Later tests used 2.00, 2.65 (IC50) and 3.00 mM (IC75).
• Main results:
  • MTT test: at 0.39 and 0.78 mM, cell viability rose to 122.1% and 119.8% of untreated cells (both p less than 0.01).
  • MTT test: 1.56 mM was not significantly different from untreated cells (93.5%).
  • MTT test: viability was 31.2% at 3.13 mM, 36.8% at 6.25 mM, 27.9% at 12.50 mM and 18.0% at 25.00 mM (all p less than 0.001 versus untreated cells).
  • Neutral red test: viability was 105.6% and 89.9% at 0.39 and 0.78 mM (not significant), 73.4% at 1.56 mM and 13.5% at 3.13 mM (both p less than 0.001). There was almost no viability at the three highest doses.
  • The IC50 (dose halving viability) was 2.65 mM based on the MTT test. The IC75 was 3.00 mM.
  • Oxidative stress index was not significantly changed at 2.00 mM. It rose almost 2.0-fold at 2.65 mM and 3.0-fold at 3.00 mM compared with control.
  • Inverted microscope: cell proliferation was seen at 0.39 and 0.78 mM, and cellular degeneration including shrinkage at 3.125 mM and above.
  • Stained acrylamide-treated cells showed condensed and kidney-shaped nuclei, shrinkage, membrane blebbing and apoptotic bodies.
  • At 2.00 mM, Bax-positive cells increased, Bcl-2 staining was barely present, and some cells had PCNA-unstained nuclei.
• The authors' conclusion: The authors concluded that in SPC212 cells acrylamide had proliferative effects at low doses and inhibiting (anti-proliferative and apoptotic) effects at relatively high doses.
• Limitations the authors state:
  • The low-dose proliferation finding rested on the MTT test and microscope images. No further proliferation assay was performed.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Mesothelioma, Mesothelium, Immunohistochemistry.

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