Changes in expression of the genes involved in sphingolipid metabolism during EMT in HBEC-12KT cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F25%3AN0000205" target="_blank" >RIV/00027162:_____/25:N0000205 - isvavai.cz</a>

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

Changes in expression of the genes involved in sphingolipid metabolism during EMT in HBEC-12KT cells

Popis výsledku v původním jazyce

Poster. Abstract in publication Czech Annual Cancer Research Meeting 2025, 24.-26.11.2025, ISBN 978-80-908348-4-2 (P10). The epithelial-to-mesenchymal transition (EMT) is a key mechanism in cancer progression, enabling cells to gain motility, resist apoptosis, and acquire stem-like properties. Understanding metabolic adaptations during EMT is essential for uncovering novel therapeutic targets. In this study, we developed a unique in vitro model of gradual EMT progression using immortalized normal human bronchial epithelial cells (HBEC-12KT) continuously exposed to the environmental carcinogen benzo[a]pyrene (BaP) for up to 12 weeks. This model captures the dynamics of EMT progression and allows simultaneous monitoring of cell phenotype, intracellular sphingolipid (SL) and glycosphingolipid (GSL) composition, and transcriptional changes in associated metabolic genes. We analyzed wild-type cells, cells at early (2 weeks), intermediate (8 weeks), and late (12 weeks) stages of BaP exposure, as well as fully transformed mesenchymal-like HBEC12KT-B1 cells. Using HPLC-MS/MS and qRT-PCR, we focused on changes involved in SL and GSL metabolism and we identified stage-specific transcriptional alterations across key genes, including SPHK2, SGPL1, ST8SIA1, A4GALT, B4GALNT1, and B3GNT5. These genes are key regulators of sphingolipid and glycosphingolipid biosynthesis and degradation pathways, and their altered expression during EMT reflects metabolic reprogramming linked to cancer cell plasticity and progression. In early EMT stage (after 2 weeks), only subtle transcriptional changes were observed, despite phenotypic alterations and shifts in lipid composition, such as increased sphingosine and gangliosides GM3 and Lc3. A longer, 8-week exposure to BaP further increased cell migratory capacity, induced epithelial-to-mesenchymal transition (EMT) markers and EMT-related transcriptional regulators (SNAI1, ZEB1 and ZEB2) and it increased intracellular sphingosine, ceramide-1-phosphate, as well as a series of GSLs (glucosylceramide, lactosylceramide, GM1a, GD3, Lc3 and Gb3). In contrast, fully transformed mesenchymal-like cells exhibited significant upregulation of genes responsible for the synthesis of bioactive lipids (e.g., sphingosine-1-phosphate) and tumor-associated GSLs (e.g., GD3, Gb3, Lc3), accompanied by corresponding shifts in their lipid profiles. Furthermore, siRNA knockdown of core EMT-related transcription factors SNAI1, ZEB1, ZEB2 indicated that they may contribute significantly to the deregulation of SL/GSL metabolism, especially in the late stages of EMT. Altogether, our data suggest that transcriptional remodeling of SL/GSL metabolism occurs predominantly during the late phase of EMT and contributes mainly to the maintenance of the mesenchymal phenotype in BaP-transformed HBEC cells. The mechanisms underlying deregulation of SL/GSL levels and functions of individual SL/GSL species deserve further attention. [This study was supported by the project of the Czech Science Foundation, No. 24-10086S.]

Název v anglickém jazyce

Changes in expression of the genes involved in sphingolipid metabolism during EMT in HBEC-12KT cells

Popis výsledku anglicky

Poster. Abstract in publication Czech Annual Cancer Research Meeting 2025, 24.-26.11.2025, ISBN 978-80-908348-4-2 (P10). The epithelial-to-mesenchymal transition (EMT) is a key mechanism in cancer progression, enabling cells to gain motility, resist apoptosis, and acquire stem-like properties. Understanding metabolic adaptations during EMT is essential for uncovering novel therapeutic targets. In this study, we developed a unique in vitro model of gradual EMT progression using immortalized normal human bronchial epithelial cells (HBEC-12KT) continuously exposed to the environmental carcinogen benzo[a]pyrene (BaP) for up to 12 weeks. This model captures the dynamics of EMT progression and allows simultaneous monitoring of cell phenotype, intracellular sphingolipid (SL) and glycosphingolipid (GSL) composition, and transcriptional changes in associated metabolic genes. We analyzed wild-type cells, cells at early (2 weeks), intermediate (8 weeks), and late (12 weeks) stages of BaP exposure, as well as fully transformed mesenchymal-like HBEC12KT-B1 cells. Using HPLC-MS/MS and qRT-PCR, we focused on changes involved in SL and GSL metabolism and we identified stage-specific transcriptional alterations across key genes, including SPHK2, SGPL1, ST8SIA1, A4GALT, B4GALNT1, and B3GNT5. These genes are key regulators of sphingolipid and glycosphingolipid biosynthesis and degradation pathways, and their altered expression during EMT reflects metabolic reprogramming linked to cancer cell plasticity and progression. In early EMT stage (after 2 weeks), only subtle transcriptional changes were observed, despite phenotypic alterations and shifts in lipid composition, such as increased sphingosine and gangliosides GM3 and Lc3. A longer, 8-week exposure to BaP further increased cell migratory capacity, induced epithelial-to-mesenchymal transition (EMT) markers and EMT-related transcriptional regulators (SNAI1, ZEB1 and ZEB2) and it increased intracellular sphingosine, ceramide-1-phosphate, as well as a series of GSLs (glucosylceramide, lactosylceramide, GM1a, GD3, Lc3 and Gb3). In contrast, fully transformed mesenchymal-like cells exhibited significant upregulation of genes responsible for the synthesis of bioactive lipids (e.g., sphingosine-1-phosphate) and tumor-associated GSLs (e.g., GD3, Gb3, Lc3), accompanied by corresponding shifts in their lipid profiles. Furthermore, siRNA knockdown of core EMT-related transcription factors SNAI1, ZEB1, ZEB2 indicated that they may contribute significantly to the deregulation of SL/GSL metabolism, especially in the late stages of EMT. Altogether, our data suggest that transcriptional remodeling of SL/GSL metabolism occurs predominantly during the late phase of EMT and contributes mainly to the maintenance of the mesenchymal phenotype in BaP-transformed HBEC cells. The mechanisms underlying deregulation of SL/GSL levels and functions of individual SL/GSL species deserve further attention. [This study was supported by the project of the Czech Science Foundation, No. 24-10086S.]

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

30108 - Toxicology

Projekt

<a href="/cs/project/GA24-10086S" target="_blank" >GA24-10086S: Dopady působení polycyklických aromatických uhlovodíků na buněčné procesy spojené se stresovou signalizací a deregulací metabolismu</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů