PARP-1在人脑胶质瘤中的表达及其抑制剂BMN-673对胶质瘤细胞治疗的增敏作用

发布时间：2018-03-09 21:46

本文选题：胶质瘤　切入点：PARP-1　出处：《中南大学》2014年博士论文　论文类型：学位论文

【摘要】：目的：本实验拟研究人脑胶质瘤组织中PARP-1mRNA的表达；观察给予替莫唑胺、PARP-1抑制剂BMN-673后胶质瘤细胞中PARP-1的表达；然后对胶质瘤U251、U87细胞系予以PARP-1抑制剂BMN-673与替莫唑胺单独用药及联合用药,通过彗星实验观察细胞的DNA损伤；通过CCK-8试验、流式细胞术观察细胞增殖抑制、细胞凋亡及细胞周期阻滞水平,以初步探讨PARP-1抑制剂BMN-673与替莫唑胺联用治疗胶质瘤的可能机制,并为其临床治疗提供基础理论依据。方法：收集中南大学湘雅医院神经外科2013年10月至2014年01月35例新鲜胶质瘤、5例新鲜正常脑组织标本,用定量RT-PCR测定其肿瘤正常组组织间的PARP-1mRNA表达差异及与胶质瘤病理分级的关系。对恶性星形胶质瘤细胞系U251、恶性胶质母细胞瘤细胞系U87予以替莫唑胺、PARP-1抑制剂BMN-673处理,并通过RT-PCR测定其PARP-1的表达情况。对胶质瘤细胞予以替莫唑胺、BMN-673单独及联合用药,通过彗星实验检测DNA损伤,CCK-8试验检测细胞生长抑制、流式细胞术检测细胞凋亡及细胞周期阻滞情况。结果：胶质瘤组织中PARP-1mRNA的表达水平比正常脑组织显著升高,具有统计学意义(P0.01)；肿瘤不同级别间PARP-1mRNA表达量存在差异,具有统计学意义(P=0.01)。对胶质瘤细胞予以替莫唑胺、BMN-673及联合用药后,PARP-1mRNA表达水平较正常培养组存在差异,具有统计学意义(P0.05)。替莫唑胺与BMN-673联合用药后,U251、U87细胞的DNA损伤加重,增殖受到明显抑制,细胞凋亡增加,细胞周期G2/M期受到阻滞。结论：1. PARP-1mRNA的表达在胶质瘤组织中明显增高,且与病理级别呈正相关；2.予以PARP-1抑制剂BMN-673并与常用化疗药物TMZ联用,胶质瘤细胞中PARP-1mRNA表达升高;3.PARP-1抑制剂BMN-673对替莫唑胺在胶质瘤细胞治疗中有增敏作用。PARP-1有望成为胶质瘤分子靶向治疗的一个治疗靶点,PARP-1抑制剂BMN-673有望成为DNA损伤类化疗药物的增敏剂。
[Abstract]:Objective: to study the expression of PARP-1mRNA in human glioma tissues and to observe the expression of PARP-1 in glioma cells after treatment with temozolomide PARP-1 inhibitor BMN-673. Then the U251 U87 cell line was treated with PARP-1 inhibitor BMN-673 and temozolidomide alone or in combination. The DNA damage was observed by comet assay, and the inhibition of cell proliferation was observed by CCK-8 assay and flow cytometry. In order to explore the possible mechanism of the combination of PARP-1 inhibitor BMN-673 and temozolamide in the treatment of glioma and provide the basic theoretical basis for its clinical treatment. Methods: 35 cases of fresh normal brain tissue were collected from October 2013 to January 2014 in Xiangya Hospital of Central South University. Quantitative RT-PCR was used to determine the difference of PARP-1mRNA expression in normal tissues and its relationship with pathological grade of glioma. The malignant astrocytoma cell line U251 and malignant glioblastoma cell line U87 were treated with temozolomide PARP-1 inhibitor BMN-673. The expression of PARP-1 in glioma cells was determined by RT-PCR. The glioma cells were treated with temozolamine BMN-673 alone or in combination. DNA damage was detected by comet assay and CCK-8 assay was used to detect the growth inhibition of glioma cells. Apoptosis and cell cycle arrest were detected by flow cytometry. Results: the expression of PARP-1mRNA in gliomas was significantly higher than that in normal brain tissues (P 0.01). There were significant differences in the expression of PARP-1mRNA between the glioma cells treated with temozolamide BMN-673 and those of the normal culture group. The DNA damage of U251 U87 cells was aggravated after the combination of temozolidomide and BMN-673. Proliferation was significantly inhibited, cell apoptosis was increased and G 2 / M phase of cell cycle was blocked. Conclusion the expression of PARP-1mRNA was significantly increased in glioma tissues, and was positively correlated with pathological grade. BMN-673, a PARP-1 inhibitor, was used in combination with TMZ, a common chemotherapeutic drug. Increased expression of PARP-1mRNA in glioma cells. 3. PARP-1 inhibitor BMN-673 can enhance the sensitivity of temozolamide in glioma cells. PARP-1 may become a therapeutic target for glioma molecular targeted therapy. PARP-1 inhibitor BMN-673 may become a kind of DNA damage. An sensitizer for a therapeutic drug.
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R739.4

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