恶性脑膜瘤细胞对HER2靶向药物敏感性的研究

发布时间：2018-05-14 04:00

本文选题：恶性脑膜瘤 + HER2　；参考：《南昌大学》2016年博士论文

【摘要】：目的:探讨HER2靶向药物对HER2阳性的人恶性脑膜瘤细胞增殖、侵袭及凋亡的影响,以期为脑膜瘤个体化靶向治疗提供实验依据,对治疗复发性及手术难以彻底切除的恶性脑膜瘤具有重要意义。方法:实验对象为经q PCR检验的HER2阳性表达的人恶性脑膜瘤细胞株IOMM-Lee和CH157-MN细胞株;其中前者为中度表达,后者为弱表达。将实验分为五组:空白对照组、IOMM-Lee细胞组、IOMM-Lee-Tra用药组、CH157-MN细胞组和CH157-MN-Tra用药组。对比HER2靶向药物(曲妥珠单抗)作用前后恶性脑膜瘤细胞在体内、外增殖、侵袭及凋亡的变化;采用MTT检测各组细胞增殖及体外药物敏感性;transwell侵袭小室法检测细胞侵袭能力;流式细胞术检测各组细胞凋亡情况;Western Blot检测凋亡相关蛋白Caspase-3、PARP在药物作用前后表达的变化;裸鼠成瘤实验,观察成瘤情况,并对比药物作用前后瘤体体积的变化。结果:1、HER2靶向药物作用后:a、IOMM-Lee-Tra组在体外的增殖力及侵袭力均较IOMM-Lee组降低,两者比较差异具有统计学意义;而CH157-MN-Tra组较CH157-MN组降低不明显;b、IOMM-Lee-Tra组细胞凋亡率较IOMM-Lee组高,两者差异具有统计学意义;CH157-MN-Tra组与CH157-MN组对比凋亡率差异无具有统计学意义;2、Western Blot检测药物作用后,PARP及Caspase-3蛋白在IOMM-Lee-Tra组表达均高于IOMM-Lee组,而HER2蛋白表达在IOMM-Lee-Tra组表达低于IOMM-Lee组,差异具有统计学意义;CH157-MN-Tra组与CH157-MN组比较,PARP、Caspase-3及HER2蛋白变化不明显,差异无统计学意义。3、裸鼠体内成瘤实验发现,IOMM-Lee-Tra组瘤体体积较IOMM-Lee组缩小,差异具有统计学意义;CH157-MN-Tra组与CH157-MN组变化不明显。结论:1、中度表达HER2的恶性脑膜瘤细胞IOMM-Lee对曲妥珠单抗具一定敏感性。而曲妥珠单抗作用于弱表达HER2的恶性脑膜瘤细胞CH157-MN的效果不明显。2、曲妥珠单抗可能是通过增加PARP、Caspase-3蛋白表达,降低HER2蛋白水平,从而对HER2阳性脑膜瘤细胞产生抑瘤作用。该实验数据可能为恶性脑膜瘤的临床个体化治疗提供实验依据。
[Abstract]:Objective: to investigate the effects of HER2 targeting drugs on proliferation, invasion and apoptosis of HER2 positive human malignant meningioma cells in order to provide experimental evidence for individualized targeted therapy of meningiomas. It is of great significance for the treatment of recurrent malignant meningioma which is difficult to be resected completely. Methods: human malignant meningioma cell line IOMM-Lee and CH157-MN cell line with positive expression of HER2 were tested by Q PCR, in which the former was moderately expressed and the latter was weakly expressed. The experiment was divided into five groups: control group, IOMM-Lee cell group, IOMM-Lee-Tra group, CH157-MN cell group and CH157-MN-Tra group. The changes of proliferation, invasion and apoptosis of malignant meningioma cells in vivo, in vitro and in vitro were compared before and after treatment with HER2, and the invasion ability of malignant meningioma cells was detected by MTT assay and drug-sensitive transwell invasive chamber assay in vitro. Flow cytometry was used to detect apoptosis in each group. Western Blot was used to detect the expression of apoptosis-related protein Caspase-3PARP before and after drug treatment, and tumorigenesis was observed in nude mice before and after treatment, and the changes of tumor volume before and after drug treatment were compared. Results the proliferation and invasiveness of the IOMM-Lee group were significantly lower than that of the IOMM-Lee group, but the apoptosis rate of the CH157-MN-Tra group was lower than that of the CH157-MN group, but the apoptosis rate of the CH157-MN-Tra group was higher than that of the IOMM-Lee group. There was no significant difference in apoptotic rate between CH157-MN-Tra group and CH157-MN group. The expression of IOMM-Lee-Tra and Caspase-3 protein in IOMM-Lee-Tra group was higher than that in IOMM-Lee group, while the expression of HER2 protein in IOMM-Lee-Tra group was lower than that in IOMM-Lee group. The difference between CH157-MN-Tra group and CH157-MN group was not significant, but there was no significant difference in Caspase-3 and HER2 protein between CH157-MN-Tra group and CH157-MN group. The tumor volume of IOMM-Lee-Tra group was smaller than that of IOMM-Lee group. There was no significant difference between CH157-MN-Tra group and CH157-MN group. Conclusion IOMM-Lee of malignant meningioma cells with moderate expression of HER2 is sensitive to trotozumab. However, the effect of trotozumab on CH157-MN of malignant meningioma cells with weak expression of HER2 was not obvious. It may be that tratuzumab decreases the level of HER2 protein by increasing the expression of PARPnCaspase-3 protein, and thus has an inhibitory effect on HER2 positive meningioma cells. The experimental data may provide experimental basis for individual treatment of malignant meningioma.
【学位授予单位】：南昌大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R739.45

【相似文献】