Single-Cell FISH Analysis Reveals Distinct Shifts in PKM Isoform Populations during Drug Resistance Acquisition

The Warburg effect, i.e., the utilization of glycolysis under aerobic conditions, is recognized as a survival advantage of cancer cells. However, how the glycolytic activity is affected during drug resistance acquisition has not been explored at single-cell resolution. Because the relative ratio of...

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Published inBiomolecules (Basel, Switzerland) Vol. 12; no. 8; p. 1082
Main Authors Kim, Seong Ho, Wi, Ji Hun, Gwak, HyeRan, Yang, Eun Gyeong, Kim, So Yeon
Format Journal Article
LanguageEnglish
Published Basel MDPI AG 06.08.2022
MDPI
Subjects
Adenosine
Aerobic conditions
Analysis
Antibodies
Cancer cells
Care and treatment
Cell survival
Colorectal cancer
Diagnosis
Drug development
Drug resistance
drug response
E coli
Enzymes
Fluorescence in situ hybridization
Gefitinib
Glycolysis
Hybridization
isoform regulation
Kinases
Metabolic response
Metabolism
Methods
mRNA
Phosphorylation
Prevention
Pyruvate kinase
Pyruvate kinase deficiency
pyruvate kinase m (pkm)
Pyruvic acid
Risk factors
single-cell analysis
Transfer RNA
South Korea
St Louis Missouri
United States > US
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Summary:The Warburg effect, i.e., the utilization of glycolysis under aerobic conditions, is recognized as a survival advantage of cancer cells. However, how the glycolytic activity is affected during drug resistance acquisition has not been explored at single-cell resolution. Because the relative ratio of the splicing isoform of pyruvate kinase M (PKM), PKM2/PKM1, can be used to estimate glycolytic activity, we utilized a single-molecule fluorescence in situ hybridization (SM-FISH) method to simultaneously quantify the mRNA levels of PKM1 and PKM2. Treatment of HCT116 cells with gefitinib (GE) resulted in two distinct populations of cells. However, as cells developed GE resistance, the GE-sensitive population with reduced PKM2 expression disappeared, and GE-resistant cells (Res) demonstrated enhanced PKM1 expression and a tightly regulated PKM2/PKM1 ratio. Our data suggest that maintaining an appropriate PKM2 level is important for cell survival upon GE treatment, whereas increased PKM1 expression becomes crucial in GE Res. This approach demonstrates the importance of single-cell-based analysis for our understanding of cancer cell metabolic responses to drugs, which could aid in the design of treatment strategies for drug-resistant cancers.
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Present address: Department of Chemistry, University of British Columbia, Kelowna, BC V1V 1V7, Canada.
ISSN:2218-273X
2218-273X
DOI:10.3390/biom12081082