Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 May 8;26(10):4472.
doi: 10.3390/ijms26104472.

Differential Effects of Sphingolipids on Cell Death and Antioxidant Defenses in Type 1 and Type 2 Endometrial Cancer Cells

Agnieszka U Błachnio-Zabielska  1 , Patrycja Sadowska  1 , Urszula Chlabicz  1 , Karolina Pogodzińska  1 , Hervé Le Stunff  2 , Piotr Laudański  3   4   5 , Jacek Szamatowicz  6 , Mariusz Kuźmicki  6
Affiliations

Differential Effects of Sphingolipids on Cell Death and Antioxidant Defenses in Type 1 and Type 2 Endometrial Cancer Cells

Agnieszka U Błachnio-Zabielska et al. Int J Mol Sci. .

Abstract

Endometrial cancer (EC) is classified into two main subtypes with distinct molecular profiles. Sphingolipids, particularly ceramide and sphingosine-1-phosphate (S1P), are crucial regulators of cell survival, apoptosis, and oxidative stress. This study examined the impact of sphingolipid metabolism in Ishikawa (type 1) and HEC-1A (type 2) EC cells following the silencing of Sptlc1 and Sptlc2, which encode subunits of serine palmitoyltransferase (SPT), a key enzyme in de novo sphingolipid synthesis. Gene silencing was confirmed by RT-PCR and Western blot, while sphingolipid levels were quantified using UHPLC/MS/MS and the sphingolipid rheostat (S1P/ceramide ratio) was calculated. Cell viability (MTT assay), cell death, ROS levels (ELISA), total antioxidant capacity (TAC), catalase and caspase-3 activity, and mitochondrial membrane potential were also assessed. The obtained data showed higher ceramide levels in Ishikawa(CON) cells and higher S1P levels in HEC-1A(CON) cells, resulting in a higher sphingolipid rheostat in HEC-1A cells. SPT knockdown reduced sphingolipid levels, increased cell viability, elevated ROS levels, and decreased cell death, particularly in Ishikawa cells. Furthermore, after gene silencing, these cells exhibited reduced catalase activity and diminished TAC, indicating an impaired redox balance. These findings reveal subtype-specific responses to disrupted sphingolipid synthesis and highlight the importance of sphingolipid homeostasis in the behavior of EC cells.

Keywords: cell death; cell viability; ceramide; endometrial cancer; sphingolipid rheostat; sphingosine-1-phosphate.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
(A) The impact of Sptlc1 gene silencing on Sptlc1 mRNA in Ishikawa and HEC-1A cells; (B) The impact of Sptlc2 gene silencing on Sptlc2 mRNA in Ishikawa and HEC-1A cells; (C) The impact of Sptlc1 gene silencing on SPT1 protein content in Ishikawa and HEC-1A cells; (D) The impact of Sptlc2 gene silencing on SPT2 protein content in Ishikawa and HEC-1A cells; Data are presented as medians (interquartile range). The difference between groups and compared by Mann–Whitney U test: ** p < 0.01, **** p < 0.0001 vs. control group; b p < 0.01, vs. Ishikawa(CON) group.
Figure 2
Figure 2
(A) The impact of Sptlc1 and Sptlc2 gene silencing on cell viability in Ishikawa and HEC-1A cells; (B) The impact of Sptlc1 and Sptlc2 gene silencing on cell death in Ishikawa and HEC-1A cells. Data are presented as medians (interquartile range). The difference between groups and compared by Mann–Whitney U test: *** p < 0.001, **** p < 0.0001 vs. control group.
Figure 3
Figure 3
(A) The impact of Sptlc1 and Sptlc2 gene silencing on ROS levels in Ishikawa and HEC-1A cells; (B) The impact of Sptlc1 and Sptlc2 gene silencing on catalase activity in Ishikawa and HEC-1A cells; (C) The impact of Sptlc1 and Sptlc2 gene silencing on TAC in Ishikawa and HEC-1A cells; (D) The impact of Sptlc1 and Sptlc2 gene silencing on caspase-3 activity in Ishikawa and HEC-1A cells; Data are presented as medians (interquartile range). The difference between groups and compared by Mann–Whitney U test: ** p < 0.01, *** p <0.001, **** p < 0.0001 vs. control group; b p < 0.01, d p < 0.0001 vs. Ishikawa(CON) group.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Sung H., Ferlay J., Siegel R.L., Laversanne M., Soerjomataram I., Jemal A., Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J. Clin. 2021;71:209–249. doi: 10.3322/caac.21660. - DOI - PubMed
    1. Bokhman J.V. Two pathogenetic types of endometrial carcinoma. Gynecol. Oncol. 1983;15:10–17. doi: 10.1016/0090-8258(83)90111-7. - DOI - PubMed
    1. Setiawan V.W., Yang H.P., Pike M.C., McCann S.E., Yu H., Xiang Y.B., Wolk A., Wentzensen N., Weiss N.S., Webb P.M., et al. Type I and II endometrial cancers: Have they different risk factors? J. Clin. Oncol. 2013;31:2607–2618. doi: 10.1200/JCO.2012.48.2596. - DOI - PMC - PubMed
    1. Hamilton C.A., Cheung M.K., Osann K., Chen L., Teng N.N., Longacre T.A., Powell M.A., Hendrickson M.R., Kapp D.S., Chan J.K. Uterine papillary serous and clear cell carcinomas predict for poorer survival compared to grade 3 endometrioid corpus cancers. Br. J. Cancer. 2006;94:642–646. doi: 10.1038/sj.bjc.6603012. - DOI - PMC - PubMed
    1. Hannun Y.A., Obeid L.M. Sphingolipids and their metabolism in physiology and disease. Nat. Rev. Mol. Cell Biol. 2018;19:175–191. doi: 10.1038/nrm.2017.107. - DOI - PMC - PubMed

MeSH terms

LinkOut - more resources