| Objective:Laryngeal cancer is a common head and neck malignant tumor, with squamous carcinoma of the majority. The larynx are mainly composed of the comprehensive treatment such as surgery supplemented by radiation and chemotherapy, but after surgery in patients with partial or total loss of laryngeal function brings great inconvenience to the life. Although radiotherapy can save laryngeal function, improve the quality of survival, laryngeal cancer radiation resistance causes the radiation effect is not ideal, how to improve the radiation sensitivity of laryngeal cancer is a big problem of the current clinical workers need to conquer. The study found that the throat cancer is associated with multiple genes. Malignant tumor growth speed, the demand for energy is greater than normal tissue, needs a large number of glucose for energy, previous studies have found that glucose transporters (GLUT-1) in head and neck squamous cell carcinomas and high expression was associated with malignant degree and poor prognosis, and may synergy PI3k/Akt pathway in laryngeal cancer radiation resistance. We found in recent research that the GLUT-1 AS ODN, LY294002, Wortmannin can inhibit GLUT-1 and PI3K/Akt signaling pathway to increase laryngeal cancer radiation sensitivity, this study we once again detect if the change of laryngeal cancer cells GLUT-1, PI3K/Akt signal pathway can improve the radiation sensitivity of laryngeal cancer by SiRNA, Apigenin interference joint specificity PI3K/Akt signaling pathway inhibitor.Methods:Hep 2 cells in experimental transfection SiRNAGLUT-1, LY294002, wortmannin, Apigenin and different radiation dose by different combination into 15 groups: CK(control check), SiRNA(803-SiRNA), Apigenin, LY294002, Wortmannin, Apigenin +SiRNA, LY294002+SiRNA, Wortmannin+SiRNA,8Gy X-ray,8Gy X-ray+Apigenin, 8Gy X-ray+LY294002,8Gy X-ray+Wortmannin,8Gy X-ray+Apigenin+SiRNA,8Gy X-ray+LY294002+SiRNA,8Gy X-ray+Wortmannin+SiRNA, each group of five, observe and record each transplanted tumor volume, weight and radiotherapy sensitization by cell cloning experiments calculation ratio (E/O), using TUNEL apoptosis detection, real-time RT-PCR detection GLUT-1 mRNA expression, Westernblotting testingthe expression of GLUT-1, PI3K, p-Akt protein of Hep-2cells in groups.Result:E/O value:Apigenin, LY294002, Wortmannin, Apigenin+SiRNA and LY294002 +SiRNA, Wortmanin+SiRNA E/O value were 1.05,0.95,0.88,0.83,1.24,0.42TUNEL detection:before and after irradiation, Apigenin combined SiRNA and joint SiRNA Wortmannin can obviously increase the laryngeal cancer cell apoptosis, and LY294002 joint SiRNA group only reflect before irradiation have obvious effect.Real-time RT-PCR detection:before irradiation, SiRNA, LY294002, Apigenin+ SiRNA and LY294002+SiRNA could significantly cut GLUT-1 mRNA expression, each group of drugs and drug combination after irradiation SiRNA and SiRNA group compared with control group no difference between the mRNA expreson of GLUT-1 drop.Westing blooting testing:before irradiation, SiRNAGLUT-1 can enhance Apigenin to inhibittheexpression of PI3K protein of laryngeal cancer cells. After irradiation, SiRNAGLUT-1 can enhance Apigenin to inhibitiGLUT-1 of laryngeal cancer cells, p-Akt protein expression. Before irradiation, SiRNAGLUT-1 can enhance LY294002 to inhibit GLUT-1 of laryngeal cancer cells, p-Akt protein expression. After irradiation, SiRNAGLUT-1 can enhance LY294002 to inhibit GLUT-1 of laryngeal cancer cells, PI3K, p-Akt protein expression. SiRNAGLUT-1 can enhance Wortmannin before and after exposure to the GLUT-1 of laryngeal cancer cells, p-Akt inhibition of protein expression. Before and after irradiation, Wortmannin no obvious inhibition of PI3K protein.Conclusion:GLUT-1 and the excessive activation of PI3K/Akt signal pathway associated with laryngeal cancer radiation resistance, SiRNA, Apigenin, LY294002, wortmannin can not increase the laryngeal cancer radiation sensitivity. |
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