雌激素受体ER-α36在胶质瘤发生及发展中的作用初探

发布时间：2018-04-27 06:27

本文选题：ER-α36 + 胶质瘤　；参考：《山东大学》2017年硕士论文

【摘要】：胶质瘤是最为常见的脑部肿瘤,约占原发性神经系统肿瘤的60%,其发病率存在明显的性别差异,男性发病率是女性的1.85倍。胶质瘤具有复发率高、治疗效果不佳,预后极差等特点,多数患者的生存期一般小于一年。目前,针对胶质瘤的治疗方法主要是手术切除,然后辅助以放疗、化疗,但是三种方法结合也仅能极其有限地延长病人的生存期。对于胶质瘤的治疗,近年来越来越多的研究小组将研究方向转向靶向药物的研究,而靶向药物的研究最根本的是寻找治疗疾病的靶标。因为胶质瘤在男女中的发病率存在明显差异,我们推测雌激素受体可能在胶质瘤的发生发展中发挥特定作用。雌激素受体-α36(estrogen receptor-α36,ER-α36)是一种新型雌激素受体,其主要定位在细胞膜上,分子质量为35.7 kD,介导由膜起始的雌激素信号通路。已证实ER-α36参与多种癌症的发生发展,其在乳腺癌中的作用已经研究得较为深入,但是ER-α36在胶质瘤发生中的作用至今不清。本研究发现在临床采集的胶质瘤样本中,肿瘤ER-α36的表达量明显高于瘤旁。下调胶质瘤细胞系中ER-α36的表达后,胶质瘤细胞的增殖、迁移、侵袭能力明显减弱。同时,荷瘤实验结果证明,敲低ER-α36后,胶质瘤细胞的成瘤能力显著降低。因此,我们推测,ER-α36和胶质瘤的发生和发展有密切关系,可能成为临床治疗胶质瘤的新靶标。背景胶质瘤是最为常见的脑部肿瘤,发病率存在明显的性别差异。ER-α36是一种新型的膜雌激素受体,其生物学作用正在引起越来越多的关注。因为胶质瘤的发病存在显著性别差异,所以我们推测激素受体可能在胶质瘤的发生及发展中发挥作用。有文献报道,ER-α36高表达显著升高乳腺癌患病风险,而且ER-α36介导的快速雌激素信号传导通路与乳腺癌干/祖细胞功能密切相关。另有文献报道,ER-α36被激活后能够刺激子宫内膜细胞的增生。有研究表明,ER-α36在胃癌、骨质疏松等疾病发生中也发挥重要作用,ER-α36的表达量增高能够削弱化疗药物对乳腺癌细胞的杀伤作用。ER-α36和多种肿瘤的发生和发展具有密切的关系,其有望成为肿瘤治疗的新靶点。虽然已经明确ER-α36参与了乳腺癌等若干种肿瘤的发生和发展,但是其在胶质瘤发生发展中的作用目前尚未见报道。目的建立ER-α36敲低的胶质瘤细胞株,用MTT实验、划痕实验和Transwell实验研究ER-α36敲低后对细胞的增殖、迁移、侵袭等造成的影响,并用荷瘤实验进行在体验证,明确ER-α36敲低后对胶质瘤细胞的增殖、迁移、侵袭等的影响。在此基础上,进一步研究介导上述作用的信号通路,初步探明胶质瘤中ER-α36发挥作用的分子机制。方法临床采集胶质瘤及瘤旁组织,利用Western blot检测ER-α36表达,观察分析ER-α36在肿瘤组织和瘤旁组织的表达情况。分别采用ER-α36激动剂G1和抑制剂IC162处理胶质瘤细胞株观察细胞增殖的改变,初步明确ER-α36在胶质瘤细胞增殖过程中的作用。将shER-α36质粒转入野生型胶质瘤细胞U87和U251中,构建ER-α36稳定敲低的细胞株,MTT实验检测细胞增殖能力的改变并通过裸鼠荷瘤实验进行在体验证,进一步明确敲低ER-α36对胶质瘤细胞增殖中的影响。通过划痕实验及Transwell等实验检测ER-α36敲低后,胶质瘤细胞迁移以及侵袭能力的改变,同时Western blot检测上皮以及间质细胞标志蛋白。随后,检测可能参与胶质瘤增殖、迁移、侵袭作用的信号通路,初步探明ER-α36参与胶质瘤增殖、迁移、侵袭等行为的可能机制。结果(1)Western blot检测结果表明,在12对胶质瘤样本中,其中10对样本的ER-α36在肿瘤组织中的表达量高于瘤旁组织。(2)MTT结果表明随着ER-α36特异性激动剂G1浓度增加,胶质瘤细胞的增殖能力增强,而ER-α36特异性抑制剂IC162可以显著降低胶质瘤细胞的增殖能力。(3)MTT检测结果证实,稳定敲低ER-α36的胶质瘤细胞,其增殖能力较对照细胞减弱。裸鼠荷瘤实验结果表明,胶质瘤细胞中ER-α36敲低后,其体内成瘤能力相较于对照组胶质瘤细胞明显降低。(4)划痕实验以及Transwell实验结果表明,ER-α36敲低能够降低胶质瘤细胞的迁移和侵袭能力。Western blot检测上皮细胞以及间质细胞标志性蛋白,ER-α36敲低能够抑制胶质瘤细胞的上皮间质转化(epithelial-mesenchymal transition,EMT)过程。(5)Western blot 结果表明,ER-α36 敲低后 MAPK/ERK 和 PI3K/AKT 信号通路的激活减弱。结论ER-α36参与了胶质瘤的增殖、迁移、侵袭等过程,并与胶质瘤的EMT过程密切相关,这些作用可能是通过MAPK/ERK和PI3K/AKT信号通路来实现的。
[Abstract]:Glioma is the most common brain tumor, accounting for about 60% of the primary nervous system tumors. The incidence of glioma has a significant sex difference, and the incidence of male is 1.85 times. The recurrence rate of the glioma is high, the treatment effect is poor, the prognosis is very poor and so on. Most of the patients are usually less than one year. At present, the treatment of glioma The main methods are surgical resection, and then adjuvant chemotherapy and radiotherapy, but the combination of the three methods can only prolong the patient's survival time. For the treatment of glioma, more and more research teams have turned the research direction to the target drug research in recent years, and the target drug research is the most important target for the target treatment of the disease. There is a significant difference in the incidence of glioma in men and women. We speculate that the estrogen receptor may play a specific role in the development of glioma. Estrogen receptor - alpha 36 (estrogen receptor- alpha 36, ER- alpha 36) is a new type of estrogen receptor, which is mainly located on the cell membrane, and the molecular mass is 35.7 kD, mediated by the membrane initiation. The estrogen signaling pathway has been confirmed that ER- alpha 36 has been involved in the development of various cancers, and its role in breast cancer has been studied in depth, but the role of ER- alpha 36 in the occurrence of glioma is not clear. This study found that the expression of ER- alpha 36 was significantly higher than that of the tumor in the clinical samples of glioma. After the expression of ER- alpha 36 in the cell line, the proliferation, migration and invasiveness of glioma cells decreased significantly. At the same time, the tumor bearing experimental results showed that the tumorigenesis ability of glioma cells decreased significantly after knocking down ER- alpha 36. Therefore, we speculate that ER- alpha 36 is closely related to the occurrence and development of glioma, and may become a new target for the clinical treatment of glioma. Background glioma is the most common brain tumor. There is a significant gender difference in the incidence of.ER- a 36 is a new type of membrane estrogen receptor. Its biological role is causing more and more attention. Because of the significant gender difference in the pathogenesis of glioma, we speculate that the hormone receptor may be in the development and development of glioma. It is reported that the high expression of ER- alpha 36 significantly increases the risk of breast cancer, and the ER- alpha 36 mediated rapid estrogen signaling pathway is closely related to the function of the stem / progenitor cells of breast cancer. It is also reported that the activation of ER- alpha 36 can stimulate the proliferation of endometrium cells. Studies have shown that ER- alpha 36 is in gastric cancer and bone thinning. It also plays an important role in the occurrence of pine and other diseases. The increase of ER- alpha 36 can weaken the killing effect of chemotherapeutic drugs on breast cancer cells..ER- alpha 36 is closely related to the occurrence and development of various tumors. It is expected to be a new target for cancer treatment. Although it has been clear that ER- alpha 36 has been involved in a number of cancers, such as breast cancer But its role in the development of glioma has not yet been reported. Objective to establish a glioma cell line of ER- alpha 36 knockout. The effects of ER- alpha 36 on the proliferation, migration and invasion of the cells were studied by MTT experiment, scratch test and Transwell test, and the tumor bearing experiment was used to identify the ER- alpha 36. On the basis of the effects of the proliferation, migration and invasion of glioma cells. On this basis, we further study the signal pathway which mediate the above action and preliminarily explore the molecular mechanism of ER- alpha 36 in glioma. Methods the clinical collection of glioma and para tumor tissue, the detection of ER- alpha 36 expression by using blot blot, and the observation and analysis of ER- alpha 36 in tumor tissue and tumor. ER- alpha 36 agonist G1 and inhibitor IC162 were used to observe the proliferation of glioma cell lines, and the role of ER- alpha 36 in the proliferation of glioma cells was preliminarily identified. ShER- alpha 36 plasmids were transferred into the wild glioma cells U87 and U251 to construct ER- alpha 36 with a stable low cell strain and MTT experimental examination. The changes in cell proliferation ability were measured and the effect of ER- alpha 36 on the proliferation of glioma cells was further clarified through the nude mice bearing experiment. The changes in the migration and invasion of glioma cells were detected by the scratch test and the Transwell and other tests. At the same time, the Western blot was used to detect the epithelium and the interstitial fine. Then, the possible mechanism of ER- alpha 36 involved in the proliferation, migration and invasion of glioma was detected. Results (1) the results of Western blot detection showed that in 12 pairs of glioma samples, the expression of ER- alpha 36 in 10 pairs of samples was high. (2) MTT results showed that the proliferation ability of glioma cells increased with the increase of ER- alpha 36 specific agonist G1 concentration, while ER- alpha 36 specific inhibitor IC162 could significantly reduce the proliferation ability of glioma cells. (3) MTT detection results confirmed that the glioma cells with low ER- alpha 36 were stable and the proliferation ability was weaker than that of the control cells. The tumor bearing test in nude mice showed that the tumorigenicity of ER- alpha 36 in the glioma cells was significantly lower than that of the control group. (4) the scratch test and the results of Transwell experiment showed that the ER- alpha 36 knock down could reduce the migration of glioma cells and the invasive energy.Western blot to detect epithelial cells and stromal cells. ER- alpha 36 knocks low to inhibit the epithelial mesenchymal transition (epithelial-mesenchymal transition, EMT) of glioma cells. (5) Western blot results show that the activation of MAPK/ERK and PI3K/AKT signaling pathway is weakened after ER- alpha 36 knocks. Conclusion ER- alpha 36 participates in the proliferation, migration, invasion and other processes of glioma and is associated with glioma. The EMT process is closely related, and these effects may be achieved through MAPK/ERK and PI3K/AKT signaling pathways.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

【相似文献】