Objective:To investigate the effect of hydroxycamptothecin on proliferation and energy metabolism of human prostate cancer PC-3 cells and on the critical enzymes in these cells. MethodsUsing MTT method, the proliferation inhibitory ability was determined after the PC-3 cells were treated with hydroxycamptothecin at 1.0,2.0 and 5.0 μmol/L for 24h. The energy metabolism parameters related to glucose uptake, lactic acid production and ATP production were determined with spectrophotometry. The expression of lactate dehydrogenase A (LDHA) and AMP-activated protein kinase (AMPK) was detected by Western blotting.  Results:Hydroxycamptothecin inhibited proliferation of the PC-3 cells at 1.0,2.0 and 5.0 μmol/L in a dose-dependent manner (P<0.05, P<0.01), with an IC50 value of 3.24 μmol/L. Compared with the control group, hydroxycamptothecin at different concentrations could reduce glucose uptake, inhibit lactic acid production and ATP production in a dose-dependent manner (P<0.05, P<0.01). Western blotting results showed that hydroxycamptothecin at different concentrations could reduce the expression of LDHA and AMPK in PC-3 cells (P<0.05, P<0.01).  Conclusion:Hydroxycamptothecin inhibits aerobic glycolysis and cell proliferation in the tumor by reducing the expression of LDHA and AMPK in PC-3 cells.