ErbB2与其自身抑制因子的作用位点及其相互作用机制的研究
发布时间：2019-06-28 09:49
【摘要】：ErbB2是原癌基因erbB-2编码的185kDa的细胞膜受体,为表皮生长因子受体(epidermal growth factor receptor,EGFR)家族成员之一。该家族包括ErbB-1(又称EGFR,HER1)、ErbB2(又称HER2)、ErbB-3(HER3)和ErbB-4(HER4)四个成员。ErbB2高表达肿瘤细胞中Ras-MAPK和PI3K-Akt信号传导活性较高,细胞增殖能力较强,分化成熟和凋亡机制受到抑制,细胞恶性程度高。ErbB2高表达肿瘤细胞可抵抗TNF-α、射线以及各种化疗药物引起的细胞凋亡效应。临床上ErbB2表达与患者预后密切相关,ErbB2高表达的患者易发生肿瘤转移,存活期短。由于ErbB2在正常细胞和肿瘤细胞中的表达水平具有显著的差异,因而已成为肿瘤免疫生物治疗的理想靶点,亦是目前肿瘤治疗研究领域的热点分子。Herstatin是近年来发现的唯一天然存在的ErbB2自身抑制分子,它是ErbB2mRNA加工过程中选择性剪接的产物,是一种可溶性的ErbB2受体。Herstatin能与细胞表面的ErbB2受体高亲和力结合,阻碍ErbB2同源或异源二聚体的形成,抑制受体酪氨酸磷酸化及ErbB2过表达细胞的增殖能力。但是,Herstatin与ErbB2相互作用的机制及与ErbB2受体分子的结合位点以及抑制ErbB2过表达细胞增殖的机制均不清楚。阐明ErbB2分子表达的自身调节机制,有助于理解该受体分子在生理条件下表达平衡对于细胞正常分化的重要意义,而且可能为ErbB2过表达肿瘤的治疗提供新思路。对二者相互作用位点的分析亦可为寻找到控制ErbB2的敏感靶位及有效的小分子抑制剂的设计提供依据。 本研究采用基因工程技术构建了Herstatin的真核表达载体,将构建成功的载体转染ErbB2表达水平不同的细胞,在mRNA和蛋白水平均检测到Herstatin的表达。应用MTT法证实了Herstatin对细胞的增殖具有明显的抑制作用。Hoechst和PI染色发现Herstatin转染后可诱导转染细胞凋亡,这可能是其抑制细胞增殖
[Abstract]:ErbB2 is a cell membrane receptor of 185kDa encoded by proto-oncogene erbB-2 and is a member of the (epidermal growth factor receptor,EGFR family. The family includes four members: ErbB-1 (also known as EGFR,HER1), ErbB2 (also known as HER2), ErbB-3 (HER3) and ErbB-4 (HER4). ErbB2 highly expressed tumor cells have high Ras-MAPK and PI3K-Akt signal transduction activity, strong cell proliferation ability, inhibition of differentiation, maturation and apoptosis, and high degree of malignancy. ErbB2 high expression tumor cells can resist TNF- 伪, radiation and apoptosis induced by various chemotherapeutic drugs. The expression of ErbB2 is closely related to the prognosis of patients. Patients with high expression of ErbB2 are prone to tumor metastasis and have a short survival period. Because the expression level of ErbB2 in normal cells and tumor cells is significantly different, it has become an ideal target of tumor immunobiotherapy and a hot molecule in the field of tumor therapy at present. Herstatin is the only natural self-inhibitory molecule of ErbB2 discovered in recent years, and it is the product of selective splicing in the process of ErbB2mRNA processing. It is a soluble ErbB2 receptor. Herstatin can bind to ErbB2 receptor on cell surface with high affinity, hinder the formation of ErbB2 homologous or heterodimer, and inhibit the phosphorylation of receptor tyrosine and the proliferation of ErbB2 overexpression cells. However, the mechanism of the interaction between Herstatin and ErbB2, the binding site of ErbB2 receptor molecule and the mechanism of inhibiting the proliferation of ErbB2 overexpression cells are not clear. To elucidate the self-regulation mechanism of ErbB2 molecule expression is helpful to understand the significance of the expression balance of ErbB2 molecule in the normal differentiation of cells under physiological conditions, and may provide a new idea for the treatment of ErbB2 overexpression tumor. The analysis of the interaction sites can also provide a basis for finding sensitive targets to control ErbB2 and the design of effective small molecular inhibitors. In this study, the eukaryotic expression vector of Herstatin was constructed by genetic engineering technique. The constructed vector was transformed into cells with different expression levels of ErbB2, and the expression of Herstatin was detected at both mRNA and protein levels. It was confirmed by MTT assay that Herstatin had obvious inhibitory effect on cell proliferation. Hoechst and PI staining showed that Herstatin could induce apoptosis of transfected cells, which may be due to its inhibition of cell proliferation.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R346
本文编号：2507201
[Abstract]:ErbB2 is a cell membrane receptor of 185kDa encoded by proto-oncogene erbB-2 and is a member of the (epidermal growth factor receptor,EGFR family. The family includes four members: ErbB-1 (also known as EGFR,HER1), ErbB2 (also known as HER2), ErbB-3 (HER3) and ErbB-4 (HER4). ErbB2 highly expressed tumor cells have high Ras-MAPK and PI3K-Akt signal transduction activity, strong cell proliferation ability, inhibition of differentiation, maturation and apoptosis, and high degree of malignancy. ErbB2 high expression tumor cells can resist TNF- 伪, radiation and apoptosis induced by various chemotherapeutic drugs. The expression of ErbB2 is closely related to the prognosis of patients. Patients with high expression of ErbB2 are prone to tumor metastasis and have a short survival period. Because the expression level of ErbB2 in normal cells and tumor cells is significantly different, it has become an ideal target of tumor immunobiotherapy and a hot molecule in the field of tumor therapy at present. Herstatin is the only natural self-inhibitory molecule of ErbB2 discovered in recent years, and it is the product of selective splicing in the process of ErbB2mRNA processing. It is a soluble ErbB2 receptor. Herstatin can bind to ErbB2 receptor on cell surface with high affinity, hinder the formation of ErbB2 homologous or heterodimer, and inhibit the phosphorylation of receptor tyrosine and the proliferation of ErbB2 overexpression cells. However, the mechanism of the interaction between Herstatin and ErbB2, the binding site of ErbB2 receptor molecule and the mechanism of inhibiting the proliferation of ErbB2 overexpression cells are not clear. To elucidate the self-regulation mechanism of ErbB2 molecule expression is helpful to understand the significance of the expression balance of ErbB2 molecule in the normal differentiation of cells under physiological conditions, and may provide a new idea for the treatment of ErbB2 overexpression tumor. The analysis of the interaction sites can also provide a basis for finding sensitive targets to control ErbB2 and the design of effective small molecular inhibitors. In this study, the eukaryotic expression vector of Herstatin was constructed by genetic engineering technique. The constructed vector was transformed into cells with different expression levels of ErbB2, and the expression of Herstatin was detected at both mRNA and protein levels. It was confirmed by MTT assay that Herstatin had obvious inhibitory effect on cell proliferation. Hoechst and PI staining showed that Herstatin could induce apoptosis of transfected cells, which may be due to its inhibition of cell proliferation.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R346
【共引文献】
相关硕士学位论文 前2条
1 李妞妞;Maspin蛋白和C-erbB-2蛋白在子宫内膜样腺癌中的表达及其临床意义[D];吉林大学;2011年
2 崔小健;Maspin和C-erbB-2基因在前列腺癌中表达及两者相关性的初步研究[D];天津医科大学;2006年
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