Kim, Seong Ho Wi, Ji Hun Gwak, HyeRan Yang, Eun Gyeong Kim, So Yeon 2024-01-12T03:00:46Z 2024-01-12T03:00:46Z 2022-08-24 2022-08 https://pubs.kist.re.kr/handle/201004/76639 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. English Multidisciplinary Digital Publishing Institute (MDPI) Single-Cell FISH Analysis Reveals Distinct Shifts in PKM Isoform Populations during Drug Resistance Acquisition Article 10.3390/biom12081082 1 Biomolecules, v.12, no.8 Biomolecules 12 8 Y scie scopus 000846098100001 Biochemistry & Molecular Biology Biochemistry & Molecular Biology Article PYRUVATE-KINASE M2 COLORECTAL-CANCER IN-SITU EXPRESSION PROMOTES GROWTH GENE HETEROGENEITY METASTASIS drug response isoform regulation pyruvate kinase m (pkm) single-cell analysis