AMPK Knockout Impairs the Formation of Three-Dimensional Spheroids

AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis, and AMPK contributes to cell growth, apoptosis, and autophagy. Although most cell studies have been performed using two-dimensional (2D) cell culture, recent studies have demonstrated that the three-dimensi...

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Published inLife (Basel, Switzerland) Vol. 15; no. 4; p. 525
Main Authors Park, Yea-In, Park, Rackhyun, Lee, Siyun, Lee, Chunghyeon, Yoo, Inkyu, Ka, Hakhyun, Huh, Yang Hoon, Hong, Jongkwang, Park, Junsoo
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 22.03.2025
MDPI
Subjects
3D culture
AMP-activated protein kinase
AMPK
Antibodies
Apoptosis
Autophagy
Cancer
Cell culture
Cell cycle
Cell metabolism
Cell research
CRISPR
Diabetes
Energy balance
Ethanol
Flow cytometry
Homeostasis
Kinases
knockout
Laboratories
lysosome
Lysosomes
Physiological aspects
Protein kinases
spheroid
Spheroids
Three dimensional flow
South Korea
United States > US
Japan
knockout
AMPK
3D culture
lysosome
spheroid
Online AccessGet full text
ISSN2075-1729
2075-1729
DOI10.3390/life15040525

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Summary:AMP-activated protein kinase (AMPK) is an important regulator of cellular energy homeostasis, and AMPK contributes to cell growth, apoptosis, and autophagy. Although most cell studies have been performed using two-dimensional (2D) cell culture, recent studies have demonstrated that the three-dimensional (3D) spheroid technique is helpful in various cell research fields, such as tumor biology, due to its resemblance to the 3D tissue structure. However, the role of AMPK in 3D spheroid formation has not been characterized clearly. This study used the AMPK knockout cell line to examine the role of AMPK in 3D spheroid formation and is the first report describing the generation of 3D spheroids using AMPK knockout cells. While control cells produced round spheroids with a similar length-to-width ratio, AMPK knockout produced an oval shape with a more significant length-to-width ratio. We demonstrate that AMPK knockout spheroids contain significantly more prominent lysosomes in each cell, indicating that autophagic flux is impaired in 3D spheroids. Finally, flow cytometry analysis showed that AMPK knockout spheroids contain more apoptotic cells than control cells. These results indicate that AMPK is required for efficient 3D spheroid formation.
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These authors contributed equally to this work.
ISSN:2075-1729
2075-1729
DOI:10.3390/life15040525