雄激素受体信号通路对神经母细胞瘤的调节作用及复方木鸡颗粒对神经母细胞瘤治疗作用的初步观察

发布时间：2018-05-11 08:17

本文选题：神经母细胞瘤 + 雄激素受体　；参考：《大连医科大学》2017年硕士论文

【摘要】：背景:神经母细胞瘤(neuroblastoma)是儿童最常见的发生在颅外的实体肿瘤。其主要起源于肾上腺,也可起源于肾上腺以外的交感神经的其他部分,包括腹膜后或胸部。迄今导致神经母细胞瘤发病的原因还不十分清楚,但是有些遗传易感性因素被发现与神经母细胞瘤的发病有关。目前关于神经母细胞瘤的许多研究都在试图寻找有效的治疗这种恶性肿瘤的方法。虽然之前的一些报道已经表明雄激素受体(AR)在神经母细胞瘤细胞中有表达,但是雄激素受体在神经母细胞瘤中的生物学作用并不完全清楚。本研究将就雄激素受体信号通路对神经母细胞瘤的调节作用进行探讨。并初步对中药制剂复方木鸡颗粒对神经母细胞瘤的作用进行观察。方法:1.Western blot蛋白印迹法检测雄激素受体(androgen receptor AR)在神经母细胞瘤细胞系(SH-SY5Y,Neuro-2a)及其他细胞系以及转染si RNA AR后的蛋白表达水平。2.MTT法检测雄激素(R1881)、雄激素受体拮抗剂(MDV3100,ARN509)对神经母细胞瘤细胞系(SH-SY5Y,Neuro-2a)增殖能力的影响;同时检测中药复方木鸡颗粒对两种不同细胞系293T(人肾上皮细胞系)和神经母细胞瘤细胞系SH-SY5Y的IC50值的差异性。3.划痕法检测雄激素(R1881)、雄激素受体拮抗剂(MDV3100,ARN509)对神经母细胞瘤细胞系(SH-SY5Y,Neuro-2a)迁移能力的影响。4.Transwell法检测不同处理对神经母细胞瘤细胞系(SH-SY5Y)侵袭能力的影响;5.3D成球实验和软琼脂集落形成实验检测不同处理对神经母细胞瘤细胞系(SH-SY5Y,Neuro-2a)细胞球体形成能力的影响;6.裸鼠体内种植神经母细胞瘤细胞系Neuro-2a(N2a),检测雄激素在体内对神经母细胞瘤的影响。结果:1.在神经母细胞瘤细胞系(SH-SY5Y,N2a)检测出雄激素受体(AR)有高表达;2.雄激素促进而雄激素受体拮抗剂抑制体外神经母细胞瘤(SH-SY5Y,N2a)的增殖;3.雄激素促进而雄激素受体拮抗剂抑制神经母细胞瘤细胞系(SH-SY5Y,N2a)的迁移能力;4.雄激素促进而雄激素受体拮抗剂抑制神经母细胞瘤细胞系(SH-SY5Y)的侵袭能力;5.雄激素促进神经母细胞瘤细胞系(SH-SY5Y,N2a)细胞球体形成而雄激素受体拮抗剂抑制细胞球体形成的能力;6.雄激素促进神经母细胞瘤在裸鼠体内(N2a)的生长。7.复方木鸡颗粒处理后的293T和SH-SY5Y的IC50值有显著学差异。结论:研究结果表明,雄激素受体信号通路可能参与了神经母细胞瘤的进展,并且这一发现可能为神经母细胞瘤患者的诊断和治疗提供新的靶点。复方木鸡颗粒在体外能抑制神经母细胞瘤(SH-SY5Y)生长,但其具体的作用机制还需要进一步的研究。
[Abstract]:Background: neuroblastoma is the most common extracranial solid tumor in children. It originates mainly from the adrenal gland, but also from other parts of the sympathetic nerve other than the adrenal gland, including retroperitoneal or thoracic. So far, the causes of neuroblastoma are not well understood, but some genetic susceptibility factors have been found to be related to the pathogenesis of neuroblastoma. Many studies on neuroblastoma are trying to find effective treatment for this malignant tumor. Although previous reports have shown that androgen receptor ARs are expressed in neuroblastoma cells, the biological role of androgen receptors in neuroblastoma is not entirely clear. This study will explore the regulatory effects of androgen receptor signaling pathway on neuroblastoma. The effect of compound Muji granules on neuroblastoma was observed. Methods: 1. Western blot Western blot assay was used to detect the protein expression of androgen receptor in neuroblastoma cell line (SH-SY5YN Neuro-2a) and other cell lines. 2. Blot assay was used to detect the expression level of androgen receptor antagonist (R1881) and androgen receptor antagonist (TMDV3100ARN509) in neuroblastoma cell line (SH-SY5YN Neuro-2a) and after transfection of si RNA AR. The proliferation ability of SH-SY5YT Neuro-2a cell line was affected by transblastoma cell line (SH-SY5YN Neuro-2a). At the same time, the difference of IC50 value between two different cell lines 293T (human renal epithelial cell line) and neuroblastoma cell line (SH-SY5Y) was detected. Effect of scratching assay on migration ability of neuroblastoma cell line SH-SY5YN Neuro-2a in vitro. 4. Transwell method to detect the effect of different treatments on the invasiveness of neuroblastoma cell line SH-SY5Y5Y The effect of different treatments on the globular formation ability of neuroblastoma cell line SH-SY5YN Neuro-2a was detected by lipid colony forming assay. Neuroblastoma cell line Neuro-2a N2a was implanted in nude mice to detect the effect of androgen on neuroblastoma in vivo. The result is 1: 1. In neuroblastoma cell line SH-SY5YN _ 2a, androgen receptor (ARA) was found to be highly expressed in SH-SY5YN _ 2a cell line. Androgen stimulates and androgen receptor antagonist inhibits the proliferation of neuroblastoma SH-SY5YPN2a in vitro. Androgen promotes androgen receptor antagonist inhibits the migration of neuroblastoma cell line SH-SY5YON2a. Androgen promotes androgen receptor antagonist inhibits invasion of neuroblastoma cell line SH-SY5Y. Androgen promotes the formation of spheroid in neuroblastoma cell line SH-SY5YN _ 2a, and androgen receptor antagonist inhibits the formation of cell sphere. Androgen promotes the growth of neuroblastoma in nude mice. The IC50 values of 293T and SH-SY5Y treated with compound Muji granules were significantly different. Conclusion: the results suggest that the androgen receptor signaling pathway may be involved in the progression of neuroblastoma, and this discovery may provide a new target for the diagnosis and treatment of neuroblastoma. Compound Muji granules can inhibit the growth of neuroblastoma SH-SY5Y in vitro, but its specific mechanism needs further study.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.4

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