In order to study the impact of nucleus accumbens-1 (NAC1) expression on survival in cancer cell and the molecular mechanism involved. NAC1 has been observed to be up-regulated in human cancer, and to play an important role in progression and recurrence of ovarian cancer. In this study, we investigated the regulatory role of NAC1 in glycolysis and the effect of the NAC1-mediated glycolysis on cancer cell survival. Cell biology, molecular biology, and pharmacology approaches and methods were used in this study. We demonstrated: 1) NAC1 is a critical regulator of cancer cell survival under hypoxic condition. Silencing of NAC1 leads to increase in the cleaved caspase-3 and cleaved PARP, and a decline in cell viability and colony formation when the cells were subjected to hypoxia; 2) NAC1 is a novel regulator of glycolysis. Depletion of NAC1 leads to an increase in the concentration of glucose in the culture medium, a decrease in the level of intercellular lactate; 3) NAC1 enhances the stability and protein level of HIF-1 alpha. Under hypoxia, NAC1 promotes cancer cell survival and glycolysis through strengthening the stability of HIF-1 alpha.