Inhibitory Effect And Mechanism Of 2-DG Combined With Temozolomide On Glioma

ObjectiveGlioma is the most common primary intracranial tumor,accounting for 60% of the entire primary intracranial tumor,which is a malignant tumor in the brain,but the specific mechanism is not very clear at present.The symptoms and signs caused by gluoma mainly depend on the function of the brain area occupied and the placeholder effect caused by the tumor size.Its growth has the characteristics of infiltration,and the growth site is special,and it is easy to relapse,surgery often can not be completely cleared,timely and effective postoperative radiation,chemotherapy and targeted treatment,can significantly prolong the survival time,improve the prognosis.Temozolomide is a second-generation alkylation agent of imidazole and tetrazine,and it is currently the first choice of anti-glioma drugs.occupied and the placeholder effect caused by the tumor size.The increased intracranial pressure is the main manifestation,such as nausea,vomiting,headache,etc,and the corresponding symptoms of nervous system damage.Its growth has the characteristics of Although it has a significant short-term anti-cancer effect,it has a poor long-term efficacy and a relatively high level of secondary drug resistance.high level of secondary drug resistance.It has been found that combination drugs can enhance the sensitivity of temozolomide to tumor cells compared with single drugs.Therefore,the combination chemotherapy based on temozolomide has become a trend in the treatment of glioma.2-DG is a glycolytic inhibitor,which affects the glycolytic pathway by inhibiting hexokinase,and can inhibit cancer growth and promote cancer apoptosis,and its anticancer mechanism is very complex.In vitro application,it can inhibit the proliferation of a variety of malignant tumor cells,such as pancreatic cancer,breast cancer,and promote tumor cell apoptosis.Combined with other chemotherapeutic drugs,it is better than single drug in inhibiting tumor cell proliferation.In addition to inhibiting glycolysis and reducing intracellular ATP,it can also affect the REDOX reaction of cells,affect the glycosylation of glycoproteins and affect the expression of transcription factors,thereby interfering with the energy uptake of tumor cells.It can activate the intracellular unfolded protein reaction and apoptosis pathway,and then induce tumor cell apoptosis.Therefore,combined use of 2-dg as radiosensitizer and chemotherapeutic sensitizer with other anti-tumor therapies for patients with malignant tumors has a good application prospect.The purpose of this study was to investigate the in vivo and in vitro experiments: 1.2.To explore the role and mechanism of 2-DG in reversing the TMZ drug resistance of glioma cells;3.To provide the most direct evidence for evaluating the role of 2-DG and TMZ in the treatment of glioma,and to provide new treatment ideas and beneficial theoretical basis for glioma.MethodsIn vitro experiment: CCK8 method was used to determine the effects of 2-DG and TMZ on the proliferation of different types of glioma cells under different concentrations,either alone or in combination;Caspase3 activity was detected to detect apoptosis;Realtime PCR and Western blot were used to determine the apoptosis.The expression of CHOP gene;Jin Zhengjun formula evaluates the combined action of 2-DG and TMZ.In vivo experiment: Rat orthotopic glioma model was established,TMZ,2-DG,and combined administration were given for two weeks.After 14 days of administration,half of the rats were sacrificed,the tumor size was determined,and the specimen was taken for pathological examination.Rats were used to observe animal survival and to perform correlation analysis.Results2-DG has a significant inhibitory effect on a variety of glioma cells in a dosedependent manner.Among them,U251 cells have the highest sensitivity to 2-DG,so U251 cells were selected for further experimental investigation.2-DG can increase the activity of Caspase3 in U251 cells and induce apoptosis.Further studies found that 2-DG can induce high expression of CHOP gene.2-DG combined with TMZ has obvious proliferation inhibition effect on U251/TR cells,and there is a synergistic effect(Q>1.15),which can significantly increase the activation of CHOP gene compared with single drug.After CHOP silencing,the synergistic effect between 2-DG and temozolomide disappeared,suggesting that the combined use of 2-DG and temozolomide via CHOP dependent apoptosis pathway had an effect on tumor cells.When combined with temozol-omide,2-DG had obvious antitumor effect and prolonged survival in rats.HE staining showed that tumor cells in the combination group were more sparse than those in the control group,apoptotic cells became smaller and rounded,and the nuclei were fragmented and pyknoti.Conclusions2-DG significantly inhibited the growth of glioma U251 cells.The synergistic effect of2-DG in combination with temozolomide is closely related to the activation of CHOP dependent apoptosis pathway.In vivo and in vitro experiments confirmed that 2-DG can increase the anti-tumor sensitization of temozolomide and reverse its drug resistance.This study provides the theoretical basis for 2-DG-assisted temozolomide in the treatment of glioma.