Drug resistance remains a serious issue of clinical importance and is a consequence of cancer stemness. In this study, we showed that the level of Aldolase A (ALDOA) expression is significantly associated with the IC50 value of chemotherapy drugs in lung cancer. Our data revealed that ALDOA overexpression resulted in a significant increase of lung tumor spheres. The use of ingenuity pathway analysis (IPA) resulted in the identification of POU5F1 (Oct4) as the leading transcription factor of ALDOA. We observed high expression of ALDOA, Oct4 and stemness markers in collected spheroid cells. DUSP4 and TRAF4 were confirmed as major downstream targets of the ALDOA-Oct4 axis. Knockdown of these molecules significantly decreased the stemness ability of cells. In addition, we investigated whether miR-145 targets the 3′-UTR of Oct4 and is regulated by ALDOA due to the involvement of ALDOA in glycolysis and metabolic reprogramming. Furthermore, we constructed several mutant forms of ALDOA that disrupted its enzymatic activity and showed that they still induced significant in vitro sphere formation and in vivo tumorigenicity. These results demonstrated that ALDOA-mediated spheroid formation is independent of its enzymatic activity. In the clinical component, we also showed that the combination of ALDOA and TRAF4 or DUSP4 is positively correlated with poor overall survival in a xenograft model and cancer patients through immunohistochemical analyses. The results of our study revealed novel functional roles of ALDOA in inducing cancer stemness via the inhibition of miR-145 expression and the activation of Oct4 transcription. These findings offer new therapeutic strategies for modulation of lung cancer stemness to enhance chemotherapeutic responses in lung cancer patients.

耐药性仍然是临床重要性的一个严重问题，并且是癌症干的结果。在这项研究中，我们显示了醛缩酶A（ALDOA）的表达水平与肺癌化疗药物的IC50值显着相关。我们的数据显示ALDOA过表达导致肺肿瘤球的显着增加。运用独创性途径分析（IPA）鉴定了POU5F1（Oct4）作为ALDOA的主要转录因子。我们观察到在收集的球状细胞中ALDOA，Oct4和干性标记物高表达。DUSP4和TRAF4被确认为ALDOA-Oct4轴的主要下游目标。击倒这些分子会大大降低细胞的干细胞能力。此外，我们研究了miR-145是否靶向Oct4的3'-UTR，并且由于ALDOA参与糖酵解和代谢重编程而受到ALDOA的调控。此外，我们构建了ALDOA的几种突变形式，这些形式破坏了其酶活性，并显示它们仍然诱导显着的体外球体形成和体内致瘤性。这些结果证明，ALDOA介导的球体形成与其酶活性无关。在临床方面，我们还显示，通过免疫组织化学分析，ALDOA和TRAF4或DUSP4的组合与异种移植模型和癌症患者的不良整体生存呈正相关。我们的研究结果揭示了ALDOA通过抑制miR-145表达和激活Oct4转录在诱导癌症干中具有新的功能作用。这些发现为调节肺癌干细胞以增强肺癌患者的化学治疗反应提供了新的治疗策略。