Antiangiogenesis therapy has shed new light on cancer treatment, but its effectiveness, especially for overall patient survival, is still controversial. Here, we show that antiangiogenesis treatment causes a persistent suppression of mitochondria biogenesis in colorectal cancer cells, which renders them more sensitive to glycolytic blockade therapy. We first analyzed bevacizumab-resistant colon cancer xenografts by two-dimensional Blue Native/SDS-PAGE and found a serious and persistent loss of mitochondrial protein complex I. Further metabolic assays revealed significantly impaired mitochondrial function and hyperactive glycolysis, which were concomitant with the upregulation of HIF-1 and Hsp70. The treatment of bevacizumab-resistant cells with the glycolysis inhibitor 3-BrPA caused cell senescence in vitro. Intraperitoneal injection of 3-BrPA to xenograft mice bearing bevacizumab-resistant cells also resulted in smaller tumor volume and longer survival. These data provide direct evidence for the mitochondrial destruction of bevacizumab-resistant tumor cells and suggest that glycolysis blockade may potentiate the therapeutic effect of antiangiogenesis treatment. Mol Cancer Ther; 12(5); 717–24. ©2013 AACR.