Anaplastic thyroid carcinoma (ATC) is the most aggressive type of thyroid malignancies and resistant to chemotherapy. Little is known on the underlying mechanisms of ATC resistance to chemotherapy. In our work, we identified that 6-phosphogluconate dehydrogenase (6PGD) is critically involved in the development of ATC resistance to doxorubicin. We found that 6PGD mRNA, protein and enzyme activity levels are significantly upregulated in ATC cells during the prolonged exposure to doxorubicin in a time dependent manner. 6PGD inhibition by genetic and pharmacological approaches significantly inhibits growth and survival of ATC cells that are highly resistant to doxorubicin. Consistently, 6PGD inhibition also sensitizes ATC cells to doxorubicin treatment. Of note, we observed the decreased level of NADPH, NADH and enzymatic activity of sirtuin-1 in response to 6PGD inhibition in doxorubicin-resistant ATC cells. Lactate level was also reduced by 6PGD inhibition. All these indicate that 6PGD inhibition disrupts metabolic reprogramming in doxorubicin-resistant ATC cells. Our work demonstrates 6PGD activation mediated resistance in response to doxorubicin and provides an alternative therapeutic strategy to overcome resistance to chemotherapy for ATC treatment. Our findings also highlight the importance of metabolic reprogramming in ATC chemoresistance. (C) 2018 Elsevier Inc. All rights reserved.