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Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models

  • Authors:
    • Gamze Demirel
    • Cagdas Kaya
    • Ranan Gulhan Aktas
  • View Affiliations / Copyright

    Affiliations: Department of Medical Biology and Genetics, Faculty of Medicine, Maltepe University, 34857 Istanbul, Türkiye, Istanbul Training and Research Hospital, Department of Internal Medicine, 34080 Istanbul, Türkiye, Cellorama, Milton, MA 02186, USA
    Copyright: © Demirel et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 27
    |
    Published online on: March 24, 2026
       https://doi.org/10.3892/mi.2026.311
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Abstract

Although there have been studies on the effect of sucrose on cell growth in various cancer types and spheroid formation, further research on whether sucrose supports the development of liver cancer cells would be instrumental for the development of more effective in vitro 3D models for drug discovery. Thus, the present study examined the effects of sucrose on the growth of liver cancer cells. The results revealed that sucrose at a concentration of 10 mM/l was associated with the increased growth of liver cancer cells and enhanced their ability to form spheroids. These findings challenge previous assumptions, as spheroid formation is associated with increased tumor aggressiveness. Furthermore, the observed increase in the levels of in alpha‑fetoprotein (a marker for liver cancer) with unaltered albumin levels (a marker for healthy liver function) suggested a potential shift towards a more malignant state. Live imaging revealed no significant differences in cell membrane behavior and the morphology of the cancer cells between the sucrose‑treated and untreated groups. On the whole, while the present study primarily focuses on phenotypic outcomes, the mechanisms underlying these observations remain to be elucidated in future studies. 
View Figures

Figure 1

Representative live images of
experimental groups at day 7. Data are presented as mean ± SD
(n=3). (A) 2D-cultured HepG2 cells under standard culture
conditions, (B) 2D-cultured HepG2 cells treated with 10 mM sucrose,
(C) 3D-cultured HepG2 spheroids, and (D) 3D-cultured HepG2
spheroids treated with 10 mM sucrose.

Figure 2

PAS-stained (A) 2D-growing HepG2 cells
under traditional culture conditions, (B) 2D-growing HepG2 cells
following treatment with 10 mM sucrose, (C) 3D-growing HepG2
spheroids, and (D) 3D-growing HepG2 spheroids after treatment with
10 mM sucrose. Panels (E-H) demonstrate the visualization of PAS
staining using ImageJ software. (I) Quantification of PAS staining
intensity. Data are presented as mean ± SD (n=3). Statistical
analysis was performed using two-way ANOVA to evaluate the effects
of culture condition (2D vs. 3D), treatment (control vs. sucrose),
and their interaction, followed by Tukey's multiple comparisons
test. A P-value <0.001 was considered to indicate a
statistically significant difference. *P=0.0523;
**P=0.0079 and ****P<0.0001.

Figure 3

Live-membrane and DAPI-stained (A)
2D-growing HepG2 cells under traditional culture conditions, (B)
2D-growing HepG2 cells following treatment with 10 mM sucrose, (C)
3D-growing HepG2 spheroids, and (D) 3D-growing HepG2 spheroids
following treatment with 10 mM sucrose. (E) Quantification of
membrane integrity. (F) Average cell count graph. (A-D) The image
on the left panel illustrates the plasma membrane, the image in the
middle panel illustrates the cell nucleus, and the image on the
right panel represents the merged image. Data are presented as the
mean ± SD (n=3). Statistical analysis was performed using two-way
ANOVA to evaluate the effects of culture condition, treatment, and
their interaction, followed by Tukey's multiple comparisons test. A
P-value <0.001 was considered to indicate a statistically
significant difference, as follows: (E) *P<0.226;
**P<0.255; ***P<0.005 and
****P<0.0001; (F) ***P<0.0002 and
****P<0.0001.

Figure 4

Representative confocal
immunofluorescence images of AFP and albumin expression in HepG2
cells cultured under two-dimensional (2D) and three-dimensional
(3D) conditions. (A) 2D-cultured HepG2 cells under standard culture
conditions and 2D-cultured HepG2 cells treated with 10 mM sucrose.
(B) 3D-cultured HepG2 spheroids and 3D-cultured HepG2 spheroids
treated with 10 mM sucrose. AFP is shown in green and albumin in
red. Images were acquired using confocal microscopy under identical
imaging conditions. (A-D) The image on the left panel illustrates
the antibody staining (AFP or albumin), the image in the middle
panel illustrates the cell nucleus, and the image on the right
panel represents the merged image. Data are presented as the mean ±
SD (n=3). AFP, alpha-fetoprotein.
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Copy and paste a formatted citation
Spandidos Publications style
Demirel G, Kaya C and Aktas RG: Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models. Med Int 6: 27, 2026.
APA
Demirel, G., Kaya, C., & Aktas, R.G. (2026). Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models. Medicine International, 6, 27. https://doi.org/10.3892/mi.2026.311
MLA
Demirel, G., Kaya, C., Aktas, R. G."Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models". Medicine International 6.3 (2026): 27.
Chicago
Demirel, G., Kaya, C., Aktas, R. G."Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models". Medicine International 6, no. 3 (2026): 27. https://doi.org/10.3892/mi.2026.311
Copy and paste a formatted citation
x
Spandidos Publications style
Demirel G, Kaya C and Aktas RG: Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models. Med Int 6: 27, 2026.
APA
Demirel, G., Kaya, C., & Aktas, R.G. (2026). Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models. Medicine International, 6, 27. https://doi.org/10.3892/mi.2026.311
MLA
Demirel, G., Kaya, C., Aktas, R. G."Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models". Medicine International 6.3 (2026): 27.
Chicago
Demirel, G., Kaya, C., Aktas, R. G."Sucrose fuels liver cancer cell growth and spheroid formation: New insights from 2D and 3D cell culture models". Medicine International 6, no. 3 (2026): 27. https://doi.org/10.3892/mi.2026.311
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