Poor prognosis in pancreatic cancer (PanCa) is partially due to chemoresistance to gemcitabine (GEM). Glucose metabolism has been revealed to contribute to the therapeutic resistance and pluripotent state of PanCa cells. However, few studies have focused on the effects of GEM on cancer cell metabolism, stemness of tumor cells, and molecular mechanisms that critically influence PanCa treatment. We demonstrate that GEM treatment induces metabolic reprogramming, reducing mitochondrial oxidation and upregulating aerobic glycolysis, and promotes stem-like behaviors in cancer cells. Inhibiting aerobic glycolysis suppresses cancer cell stemness and strengthens GEM’s cytotoxicity. GEM-induced metabolic reprogramming is KRAS dependent, as knockdown of KRAS reverses the metabolic shift. GEM-induced metabolic reprogramming also activates AMP-activated protein kinase (AMPK), which promotes glycolytic flux and cancer stemness. In addition, GEM-induced reactive oxygen species (ROS) activate the KRAS/AMPK pathway. This effect was validated by introducing exogenous hydrogen peroxide (H₂O₂). Taken together, these findings reveal a counterproductive GEM effect during PanCa treatment. Regulating cellular redox, targeting KRAS/AMPK signaling, or reversing metabolic reprogramming might be effective approaches to eliminate cancer stem cells (CSCs) and enhance chemosensitivity to GEM to improve the prognosis of PanCa patients.

胰腺癌（PanCa）的不良预后部分归因于对吉西他滨（GEM）的化学耐药性。葡萄糖代谢已被发现有助于PanCa细胞的治疗抗性和多能状态。但是，很少有研究集中在GEM对癌细胞代谢，肿瘤细胞干性以及严重影响PanCa治疗的分子机制的影响上。我们证明GEM治疗诱导代谢重编程，减少线粒体氧化和上调有氧糖酵解，并促进癌细胞中的茎样行为。抑制有氧糖酵解可抑制癌细胞干细胞并增强GEM的细胞毒性。GEM诱导的代谢重编程是KRAS依赖的，因为KRAS的敲除逆转新陈代谢的转变。GEM诱导的代谢重编程也可以激活AMP激活的蛋白激酶（AMPK），从而促进糖酵解通量和癌症干性。此外，GEM诱导的活性氧（ROS）激活KRAS / AMPK途径。通过引入外源过氧化氢（H ₂ O ₂）验证了该效果。综上所述，这些发现揭示了在PanCa治疗过程中产生适得其反的GEM效应。调节细胞氧化还原，靶向KRAS / AMPK信号或逆转代谢重编程可能是消除癌症干细胞（CSC）和增强对GEM的化学敏感性以改善PanCa患者预后的有效方法。