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目的探讨BRAF抑制剂PLX4032联合放疗对高级别神经胶质瘤的疗效。方法选择两株细胞分别为AM-38(BRAF V600E)和GBM8(BRAF wildtype),用Cell Titre Glo方法检测细胞的IC50;用克隆形成实验检测PLX4032对不同放射剂量的增敏作用;用流式细胞仪检测PLX4032和放疗对细胞凋亡和周期的作用。结果 PLX4032显著抑制BRAF V600E细胞的增殖(IC_(50)=2μM)而对野生型细胞没有作用;克隆形成实验发现在BRAF V600E细胞上随着剂量增加,联合治疗组比单独放疗组的抑制作用更显著(P<0.05),而野生型细胞株上联合组和单独放疗组没有显著性差异(P>0.05);流式细胞分析发现在BRAF V600E突变细胞上,联合治疗组细胞凋亡数显著增加,与对照组和单独治疗组相比有显著差异(P<0.05),而在野生型细胞株上没有显著性差异;流式分析发现在BRAF V600E突变细胞上,与对照组相比,单独放疗组和联合治疗组在G0/G1期细胞数都减少,G2/M期细胞数增加,而且联合治疗组与单独放疗组相比有显著差异。结论 BRAF抑制剂PLX4032与放疗联合治疗BRAF V600E突变的高级别神经胶质瘤的疗效较好。
Objective To investigate the effect of BRAF inhibitor PLX4032 combined with radiotherapy on high-grade gliomas. METHODS: Two cell lines, BRAF V600E and BRAF wildtype, were selected. The IC50 of cells was determined by Cell Titre Glo assay. The sensitizing effect of PLX4032 to different radiation doses was detected by clonogenic assay. The effect of PLX4032 and radiotherapy on apoptosis and cycle was examined. Results PLX4032 significantly inhibited the proliferation of BRAF V600E cells (IC 50 = 2μM) and had no effect on wild-type cells. Clonogenic assay showed that the inhibitory effect of combined treatment group was more than that of radiotherapy alone group on BRAF V600E cells (P <0.05). However, there was no significant difference between the wild-type cells and the radiotherapy alone group (P> 0.05). Flow cytometry analysis showed that the number of apoptotic cells in the BRAF V600E mutant cells was significantly increased (P <0.05), but no significant difference in the wild-type cell lines; flow cytometry analysis found BRAF V600E mutant cells, compared with the control group, radiotherapy alone The number of cells in G0 / G1 phase decreased and the number of G2 / M phase cells increased in combination group and combination therapy group, and there was significant difference between combined therapy group and radiotherapy alone group. Conclusion The combination of BRAF inhibitor PLX4032 and radiotherapy in the treatment of high-grade gliomas with BRAF V600E mutation is effective.