In Vitro And In Vivo Targeted Inhibition Of HK-Ⅱ Reversed The Warburg Effect To Improve The Radiosensitivity Of Laryngeal Carcinoma

[Objective]The relationship between the expression of Hexokinase-Ⅱ(HK-Ⅱ)and the radioresistance of laryngeal carcinoma was analyzed by detecting the expression of HK-Ⅱ in Tu212 cells in vitro and in vivo,and whether inhibiting the expression of HK-Ⅱ by HK-Ⅱsmall interfering RNA(HK-Ⅱ siRNA)can improve the radiosensitivity of laryngeal carcinoma by changing the Warburg effect.[Methods]1.Observe human laryngeal carcinoma tissue specimens,and detect the expression of HK-Ⅱ in laryngeal carcinoma tissues and adjacent tissues by immunohistochemical method.2.To observe the effect of HK-Ⅱ siRNA on the radioresistance of Tu212 cells.Cells were irradiated using a 6-MV linear accelerator.The cell viability,cell survival,proliferation,apoptosis and cell cycle of Tu212 cells were detected by trypan blue staining,colony formation assay,CCK-8 assay and flow cytometry,respectively.3.To observe the effect of HK-Ⅱ siRNA on the glycolysis of Tu212 cells after radiotherapy.The glucose consumption and lactate production of Tu212 cells were examined,and the expression of glycolysis-related metabolic enzymes in Tu212 cells were detected by qRT-PCR and Western Blot(WB).4.The effect of HK-Ⅱ siRNA on Tricarboxylic Acid Cycle(TCA)in Tu212 cells after radiotherapy was observed.The oxygen consumption and adenosine triphosphate(ATP)content of Tu212 cells were examined,and the expression of related metabolic enzymes involved in TCA cycle of Tu212 cells were detected by qRT-PCR and WB.[Results]1.The expression of HK-Ⅱ in laryngeal carcinoma tissues was higher than that in adjacent tissues by immunohistochemical method(p<0.01).2.Compared with treatment alone,HK-Ⅱ siRNA combined with X-ray treatment significantly reduced cell viability,inhibited cell survival and proliferation,and increased cell apoptosis and G2 phase arrest(all,p<0.01).3.HK-Ⅱ siRNA inhibited the production of glucose and lactate in Tu212 cells,and significantly inhibited the up-regulation of HK-Ⅱ,pyruvate kinase M2(PKM2),phosphofructokinase platelet(PFKP),lactate dehydrogenase(LD)(all,p<0.01).4.HK-ⅡsiRNA increased oxygen consumption and ATP production in Tu212 cells,and significantly inhibited the up-regulation of pyruvate dehydrogenase(PDH)and citrate synthase(CS)(all,p<0.01).[Conclusion]The inhibition of HK-Ⅱ by siRNA enhances the radiosensitivity of laryngeal carcinoma Tu212 cells by inhibiting glycolysis and partially inhibiting oxidative phosphorylation.