Chemical and Biological Integrative Research Center, Biomedical Research Division, Korea Institute of Science and Technology, Seoul 02792, Korea.
Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02792, Korea.
Biomolecules. 2022 Aug 6;12(8):1082. doi: 10.3390/biom12081082.
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.
瓦博格效应，即在有氧条件下利用糖酵解，被认为是癌细胞的生存优势。然而，在单细胞分辨率下，尚未探索到在获得耐药性过程中糖酵解活性如何受到影响。由于丙酮酸激酶 M（PKM）的剪接异构体，即 PKM2/PKM1 的相对比例可用于估计糖酵解活性，因此我们利用单分子荧光原位杂交（SM-FISH）方法同时定量 PKM1 和 PKM2 的 mRNA 水平。用吉非替尼（GE）处理 HCT116 细胞导致出现两种不同的细胞群。然而，随着细胞产生 GE 耐药性，表达降低的 PKM2 的 GE 敏感细胞群消失，而 GE 耐药细胞（Res）表现出增强的 PKM1 表达和严格调控的 PKM2/PKM1 比值。我们的数据表明，在 GE 处理时维持适当的 PKM2 水平对于细胞存活很重要，而在 GE Res 中增加 PKM1 表达变得至关重要。这种方法证明了单细胞分析对于理解癌细胞对药物的代谢反应的重要性，这可能有助于设计针对耐药性癌症的治疗策略。
