Department of Pharmacology, Toxicology & Neuroscience, LSU Health Sciences Center-Shreveport, Shreveport, LA 71130, USA.
Cancer Prev Res (Phila). 2011 Sep;4(9):1476-84. doi: 10.1158/1940-6207.CAPR-11-0028. Epub 2011 Jun 14.
Differentiated cells primarily metabolize glucose for energy via the tricarboxylic acid cycle and oxidative phosphorylation, but cancer cells thrive on a different mechanism to produce energy, characterized as the Warburg effect, which describes the increased dependence on aerobic glycolysis. The M2 isoform of pyruvate kinase (PKM2), which is responsible for catalyzing the final step of aerobic glycolysis, is highly expressed in cancer cells and may contribute to the Warburg effect. However, whether PKM2 plays a contributing role during early cancer development is unclear. In our studies, we have made an attempt to elucidate the effects of varying mitochondrial respiration substrates on skin cell transformation and expression of PKM2. Tumorigenicity in murine skin epidermal JB6 P+ (promotable) cells was measured in a soft agar assay using 12-O-tetradecanoylphorbol-13-acetate (TPA) as a tumor promoter. We observed a significant reduction in cell transformation upon pretreatment with the mitochondrial respiration substrate succinate or malate/pyruvate. We observed that increased expression and activity of PKM2 in TPA-treated JB6 P+ cells and pretreatment with succinate or malate/pyruvate suppressed the effects. In addition, TPA treatment also induced PKM2 whereas PKM1 expression was suppressed in mouse skin epidermal tissues in vivo. In comparison with JB6 P+ cells, the nonpromotable JB6 P- cells showed no increase in PKM2 expression or activity upon TPA treatment. Knockdown of PKM2 using a siRNA approach significantly reduced skin cell transformation. Thus, our results suggest that PKM2 activation could be an early event and play a contributing role in skin tumorigenesis.
分化细胞主要通过三羧酸循环和氧化磷酸化代谢葡萄糖来获取能量,但癌细胞则依赖于一种不同的机制来产生能量,这种机制被称为瓦博格效应(Warburg effect),其描述了有氧糖酵解的依赖性增加。丙酮酸激酶(PKM2)的 M2 同工型负责催化有氧糖酵解的最后一步,在癌细胞中高度表达,可能有助于瓦博格效应。然而,PKM2 是否在癌症早期发展中起作用尚不清楚。在我们的研究中,我们试图阐明不同线粒体呼吸底物对皮肤细胞转化和 PKM2 表达的影响。使用肿瘤促进剂 12-O-十四烷酰佛波醇-13-醋酸酯(TPA),通过软琼脂测定法测量了鼠皮肤表皮 JB6 P+(可促进)细胞中的肿瘤发生能力。我们观察到在用线粒体呼吸底物琥珀酸或苹果酸/丙酮酸预处理后,细胞转化明显减少。我们观察到在 TPA 处理的 JB6 P+细胞中 PKM2 的表达和活性增加,而用琥珀酸或苹果酸/丙酮酸预处理则抑制了这种作用。此外,TPA 处理还诱导了 PKM2 的表达,而体内小鼠皮肤表皮组织中 PKM1 的表达受到抑制。与 JB6 P+细胞相比,非可促进的 JB6 P-细胞在 TPA 处理后 PKM2 的表达或活性没有增加。使用 siRNA 方法敲低 PKM2 可显著降低皮肤细胞转化。因此,我们的结果表明,PKM2 的激活可能是早期事件,并在皮肤肿瘤发生中起作用。