FGF-2抵抗内质网应激诱导的细胞凋亡的机制研究

发布时间：2018-05-28 13:19

本文选题：内质网应激 + 细胞凋亡　；参考：《华中科技大学》2010年博士论文

【摘要】：随着环境的恶化和社会压力的增加,癌症、精神类疾病和神经退行性疾病成为危害人类健康的主要疾病,在对这些疾病的研究中,我们虽然取得了很大的进展,但是仍然有许多问题亟待解决,所以与癌症、精神类疾病和神经退行性疾病密切相关的细胞凋亡与抗凋亡的研究一直都是领域内研究的热点。在以往的研究中已经发现FGF-2是一种重要的多效因子,在相当多的研究报告都阐明了FGF-2与肿瘤和癌症细胞,特别是肝癌细胞的增殖、迁移,以及肿瘤和癌症细胞对化学药物和放射性治疗的抵抗作用有极为密切的关系。而随着对内质网应激研究的不断深入,研究也已经表明了一些与人类密切相关的疾病,神经退行性疾病(老年痴呆症、帕金森症),糖尿病,特别是癌症的发生、发展都与内质网应激有着密切的关系,其中一些机制已经得到了阐明,但是目前对真核细胞内质网应激的认识仍然有许多不清楚和有待解释的地方。针对FGF-2和内质网应激的研究成果都显示它们都与癌症和神经退行性疾病密切相关,那么FGF-2与内质网应激之间的关系到底是怎么样,其中的机制如何?以有的文献中没有深入的报道。我们的研究的目的就着眼于揭示FGF-2和内质网应激之间的关系,初步厘清其中的机制。本研究在前人研究的基础之上,着重研究了FGF-2(basic FGF)对于化学药物在肿瘤细胞上引起的通过内质网应激产生的凋亡的影响。本研究选择了一个常用的肿瘤细胞系肝细胞癌细胞系HepG2和人乳腺癌细胞系MCF-7作为研究材料,采用衣霉素(Tunicamycin/Tm)和毒胡萝卜素(Thapsigargin/Tg)来诱导其产生内质网应激,我们采用了放射性同位素摄入法、特异性的荧光染料染色和流式细胞法来检查各种不同的状态下HepG2和MCF-7细胞的情况,我们发现了FGF-2对肿瘤细胞HepG2和MCF-7的保护效应,我们采用了western blot的方法,检测了一系列与内质网应激相关的信号通路和关键蛋白的表达量和磷酸化水平变化,初步说明了这一保护效应相应的作用机理。我们还构建关键蛋白的高表达稳定细胞系,在此基础上检测了相关信号通路的变化,我们还采用了SiRNA技术对信号通路上的关键蛋白进行了know-down操作,由稳定细胞系以及SiRNA的结果我们确定了FGF-2对肿瘤细胞HepG2的保护效应中的关键蛋白,我们的研究展现了内质网应激反应信号通路与FGF-2之间的存在对话,通过对这种机制和细节的研究从而从一个方面阐明了内质网应激和FGF-2与肿瘤和疾病发生有着密切的关系,对于揭示癌症的发生发展过程的机理以及治疗有着重要的意义。
[Abstract]:With the deterioration of the environment and the increase of social pressure, cancer, mental illness and neurodegenerative diseases have become the main diseases that endanger human health, and we have made great progress in the research of these diseases. However, there are still many problems to be solved. Therefore, apoptosis and anti-apoptosis, which are closely related to cancer, mental diseases and neurodegenerative diseases, have always been the focus of research in the field. In previous studies, FGF-2 has been found to be an important multi-effect factor. In many studies, it has been demonstrated that FGF-2 is associated with the proliferation and migration of tumor and cancer cells, especially hepatoma cells. And the resistance of tumor and cancer cells to chemical drugs and radiation therapy is very close. And as the study of endoplasmic reticulum stress continues, studies have also shown that some diseases are closely related to humans, neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, diabetes, especially cancer). Development is closely related to endoplasmic reticulum stress, and some mechanisms have been elucidated. However, there are still many unclear and unexplained aspects about eukaryotic endoplasmic reticulum stress. Both FGF-2 and endoplasmic reticulum stress have been shown to be closely related to cancer and neurodegenerative diseases, so what is the relationship between FGF-2 and endoplasmic reticulum stress, and what is the mechanism? Some of the literature has not been reported in depth. Our aim is to clarify the mechanism of FGF-2 and endoplasmic reticulum stress. Based on previous studies, this study focused on the effects of FGF-2(basic on apoptosis induced by endoplasmic reticulum stress in tumor cells induced by chemical drugs. In this study, a commonly used tumor cell line HepG2 and human breast cancer cell line MCF-7 were used to induce endoplasmic reticulum stress. We used radioisotope uptake, specific fluorescent dye staining and flow cytometry to examine HepG2 and MCF-7 cells in different states, and we found the protective effects of FGF-2 on HepG2 and MCF-7 in tumor cells. The expression and phosphorylation level of a series of signal pathways and key proteins related to endoplasmic reticulum stress were detected by western blot, and the mechanism of this protective effect was preliminarily explained. We also constructed a stable cell line with high expression of key proteins, and then detected the changes of related signal pathways. We also used SiRNA technology to perform know-down manipulation of key proteins in signal pathways. Based on the results of stable cell lines and SiRNA, we have identified the key proteins involved in the protective effect of FGF-2 on HepG2 in tumor cells. Our studies have demonstrated the existence of a dialogue between the endoplasmic reticulum stress response signaling pathway and FGF-2. Through the study of this mechanism and the details, it is shown that endoplasmic reticulum stress and FGF-2 are closely related to tumor and disease, which is of great significance to reveal the mechanism and treatment of cancer.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R363;Q255

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