Effect of combined therapy of radiosurgery and temozolomide on U87-epidermal growth factor receptor and variant Ⅲ cells and their xenograft
10.3760/cma.j.issn.1671-8925.2019.12.005
- VernacularTitle:放射联合替莫唑胺治疗对U87-EGFRv Ⅲ细胞及移植瘤影响的实验研究
- Author:
Yiguang LIN
1
;
Dong LIU
;
Xiaomin LIU
;
Desheng XU
Author Information
1. 天津医科大学第二医院神经外科伽玛刀中心
- Keywords:
Epidermal growth factor receptor and variant Ⅲ;
Mutation;
Radiotherapy;
Temozolomide;
Invasion;
Proliferation
- From:
Chinese Journal of Neuromedicine
2019;18(12):1215-1223
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of radiosurgery in combination of temozolomide therapy in vitro and in vivo on invasion and endocrine function of human glioma cell line U87-epidermal growth factor receptor and variant Ⅲ (EGFRvⅢ) and their xenografl.Methods (1) Human glioma U87-EGFRvⅢ cells were routinely cultured;CCK-8 was used to detect the effect of temozolomide at different concentrations on proliferation of U87-EGFRvⅢ cells,and the semi-inhibitory concentration (IC50) oftemozolomide was calculated;the U87-EGFRvⅢ cells were divided into control group,6 Gy radiotherapy group and 12 Gy radiotherapy group,and the expressions of invasion-related proteins and gene repair proteins in the three groups were detected by Western blotting after 0,6 and 12 Gy edge-dose irradiation,respectively;the U87-EGFRvⅢ cells were divided into control group,6 Gy radiotherapy group and 6 Gy radiation combined with temozolomide treatment group (5 mmol/L temozolomide was added after 6 Gy irradiation),and 24 h after each treatment,Western blotting was used to detect the expressions of invasion related proteins and gene repair related proteins in the three groups.(2) Nude mouse transplantation intracranial glioma models were established with cell suspension of the control group,6 Gy radiotherapy group and 6 Gy radiation combined with temozolomide treatment group;7,14,21 d after transplantation,biolumineseence imaging (BLI) was employed to detect luciferase expression in nude micein vivo and measurement of transplanted tumors was performed;survival curve and body mass curve were drew in nude mice;immunohistochemical staining was used to detect the expressions of EGFRvⅢ,epidermal growth factor receptor (EGFR),matrix metalloproteinase (MMP)-9,vascular endothelial growth factor (VEGF) and Ki67 in intracranial transplanted gliomas.Results (1) The IC50 oftemozolomide on U87-EGFRvⅢ cells was 5 mmol/L;as compared with those in the control group,the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 significantly increased in the cells of the 6 Gy radiotherapy group,while the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 significantly decreased in the cells of the 12 Gy radiotherapy group as compared with those in the 6 Gy radiotherapy group (P＜0.05);as compared with those in the 6 Gy radiotherapy group,the expressions of EGFRvⅢ,EGFR,MMP-2,MMP-9,VEGF,MGMT,AKT-1,β-catenin and Ku70 in cells of the 6 Gy radiotherapy combined with temozolomide group were statistically decreased (P＜0.05).(2) As compared with mice in the control group and 6 Gy radiotherapy group,mice in the 6 Gy radiotherapy combined with temozolomide group had significantly lower cell tumorigenicity,significantly higher survival rate and body mass 21 d after transplantation,and significantly smaller volume of xenograft tumors (P＜0.05);the expressions of EGFRvⅢ,EGFR,MMP-9,VEGF and Ki67 in tumor tissues of nude mice in the 6 Gy radiotherapy group were significantly decreased as compared with those in the control group;as compared with those in the 6 Gy radiotherapy group,the expressions of EGFRvⅢ,EGFR,MMP-9,VEGF and Ki67 in tumor tissues of mice in the 6 Gy radiotherapy group were significantly decreased (P＜0.05).Conclusion The combined treatment of radiosurgery and temozolomide can more obviously inhibit the invasiveness of U87-EGFRvⅢ cells and it exerts a stronger inhibitory effect on tumorigenicity of U87-EGFRvⅢ cells in brain of nude rats.
