Bruceine D Inhibits Energy Metabolism In Human Triplenegative Breast Cancer MDA-MB-231 Cells

Objective: The purpose of this study was to study the effect and possible mechanism of Bruceine D on the energy metabolism of human triple-negative breast cancer MDA-MB-231 cells under in vitro culture conditions.Methods: First,the MTT method was used to detect the effect of Bruceine D on MDA-MB-231 breast cancer cell viability,and cisplatin was used as a positive control for cell viability determination.Next,we examined the effect of Bruceine D on the aerobic glycolysis process of MDA-MB-231 breast cancer cells.The glucose consumption was measured by the glucose kit,the lactic acid production was measured by the lactic acid kit,and the ATP content was measured by the ATP kit.Next,we examined the effect of Bruceine D on the activity of aerobic glycolysis key enzymes(hexokinase,6-phosphofructokinase-1,pyruvate kinase,lactate dehydrogenase)of breast cancer cells.Measure hexokinase activity by hexokinase kit,measure 6-phosphofructokinase-1 activity by phosphofructokinase kit,measure pyruvate kinase activity by pyruvate kinase kit,and measure lactate dehydrogenase activity by lactate dehydrogenase kit.Finally,Western blotting was used to detect the effect of Bruceine D on PI3 K and Akt signaling protein expression levels in MDA-MB-231 breast cancer cells,and ?-actin was used as the internal reference for Western blotting detection.Results: MTT results showed that Bruceine D inhibited cell viability in a time-and dosedependent manner,with IC50 values of 41.44,7.07,and 2.42 ?M at 24,48,and 72 hours,respectively.The measurement results of energy substances showed that the ATP content of different concentrations of Bruceine D(1,2,4?M)after 24 hours of treatment was 1.71�0.04,1.43�0.04 and 1.24�0.03?M,which were significantly lower than the control group(1.94 �0.01?M).After 24 hours of treatment with different concentrations of Bruceine D(1,2,4 ?M),the glucose consumption was 15.44 � 0.56,10.35 � 0.24 and 7.11 � 0.16 mM,which were significantly lower than the control group(17.60 � 0.18 mM).After 24 hours of treatment with different concentrations of Bruceine D(1,2,4?M),the amount of lactic acid produced was 20.98�0.42,14.87�0.39 and 10.48�0.45 mM,which was also significantly lower than the control group(23.53�0.28mM).The detection results of the key enzyme kits showed that the activities of the four key enzymes for aerobic glycolysis in MDA-MB-231 breast cancer cells were inhibited by Bruceine D to varying degrees.Western blot results showed that Bruceine D significantly down-regulated the expression of PI3 K and p-Akt,but had no significant effect on the expression of Akt.Conclusion: Bruceine D may inhibit the aerobic glycolysis process of human breast cancer MDA-MB-231 breast cancer cells and the energy supply in the cells may also be greatly reduced.This inhibitory effect has little to do with the antiproliferative activity of Bruceine D,but may be closely related to the inhibition of the PI3K/Akt signaling pathway by Bruceine D.