Recombinant Maspin Protein Inhibits the Tumor Angiogenesis by Regulating SIRT3-Mediated Glycolysis in Cervical Cancer / 中国药学杂志

ABSTRACT

OBJECTIVE: To investigate the effect of recombinant mammary serine protease inhibitor (Maspin) on tumor angiogenesis and tumor metabolism of HeLa cervical cancer and its mechanisms. METHODS: MTS was used to test the effect of recombinant Maspin on proliferation of HeLa cells. Western blotting was used to determine the effect of recombinant Maspin on the expression of SIRT3 in HeLa cells. SIRT3 siRNA was constructed by commercial and then transfected into HeLa cells. Tube formation assay was used to study the effect of SIRT3 intervene on tumor angiogenesis. Detection kits were used to examine the effects of recombinant Maspin on the glucose consumption and the generation of pyruvic acid, lactic acid as well as ATP after SIRT3 intervene, respectively. Western blotting was used to study the effect of SIRT3 intervene on expression of glucose transporter 1 (GLUT1), monocarboxylate transporter-1 (MCT-1), hexokinase (HK2), fructokinase 6-phosphate(PFK1) and pyruvic acid 2 (PKM2). RESULTS: Compared with the control group, the proliferation of HeLa cervical cancer cells was significant inhibited by recombinant Maspin in vitro. SIRT3 siRNA transfection could significantly decrease the inhibition effect of recombinant Maspin on tumor angiogenesis. SIRT3 siRNA transfection could significantly ameliorate the inhibition effect of recombinant Maspin on glucose consumption and the contents of pyruvic acid, lactic acid as well as ATP, respectively. Recombinant Maspin inhibited the expressions of GLUT1, MCT-1, HK2, PFK1 and PKM2, however, SIRT3 siRNA transfection significantly reversed the expression of above glycolysis-related proteins. CONCLUSION: Recombinant Maspin inhibits the tumor angiogenesis by suppressing glycolysis in HeLa cells, and such effect is dependent on SIRT3 expression.