IGF1R酪氨酸激酶细胞模型构建及药物筛选的应用

发布时间：2018-03-20 03:34

本文选题：IGF1R激酶受体　切入点：BaF3细胞　出处：《华东师范大学》2011年硕士论文　论文类型：学位论文

【摘要】：胰岛素样生长因子1受体(insulin-like growth factor 1 receptor, IGF1R)是酪氨酸蛋白激酶家族的成员,在各种类型细胞上均有表达。IGF1R具有潜在的有丝分裂原作用,可促进细胞的增殖、调节细胞的恶性转化、保护肿瘤细胞免受凋亡等生物功能,其信号转导途径与肿瘤的发生密切相关,因此IGF1R被认为是非常具有潜力的抗肿瘤药物的靶点。本论文构建酪氨酸激酶IGF1R组成型高表达的细胞模型,并以该细胞模型对天然产物化合物库进行高通量药物筛选,筛选出对酪氨酸激酶IGF1R具有特异性抑制作用的组分。该结果进一步验证该IGF1R酪氨酸激酶细胞模型的稳定性、可靠性及其于药物筛选的应用。 1 IGF1R酪氨酸激酶细胞模型构建以实验室已构建的哺乳动物细胞表达载体pcDNA3.1(+)-Tel为基础,将IGF1R基因与之连接构建重组质粒。将重组质粒转入IL3依赖性的小鼠原B细胞(BaF3细胞)并在G418及IL3非依赖性的条件下筛选出能够组成型高表达IGF1R激酶活性的细胞株BaF3/Tel-IGF1R.通过分别从分子水平,蛋白水平及功能水平的鉴定,说明筛选所得到BaF3/Tel-IGF1R细胞株能够正确的表达配体非依赖性的具有组成型活性的酪氨酸激酶IGF1R,为后续药物筛选的应用打下良好的基础。 2酪氨酸激酶IGF1R细胞模型的药物筛选应用以构建的BaF3/Tel-IGF1R细胞株为高通量筛选的细胞模型,从天然产物库中筛选酪氨酸激酶IGF1R的抑制组分。对于高通量筛选的结果进行初步分析,并确定有活性的组分进一步验证。通过验证确定天然产物库中酪氨酸激酶IGF1R抑制组分,为以后其特异性抑制剂的筛选提供指导意义,进而为抗肿瘤药物的开发与研究提供先导化合物。
[Abstract]:Insulin-like growth factor 1 receptor (IGF1R) is a member of tyrosine protein kinase family. To protect tumor cells from apoptosis and other biological functions, its signal transduction pathway is closely related to the occurrence of tumor. Therefore, IGF1R is considered to be a potential target of antitumor drugs. In this paper, a cell model with high expression of tyrosine kinase IGF1R was constructed, and the natural product compound library was screened by the cell model. The results showed that the IGF1R tyrosine kinase cell model was stable, reliable and useful for drug screening. Construction of 1 IGF1R tyrosine kinase cell model. Based on the mammalian cell expression vector pcDNA3.1, constructed in our laboratory, The recombinant plasmid was constructed by ligating the IGF1R gene with the recombinant plasmid. The recombinant plasmid was transferred into the IL3 dependent mouse proto-B cell line (Baf3 cell) and the cell lines with high expression of IGF1R kinase activity were screened under G418 and IL3 independent conditions. BaF3 / Tel-IGF1R. The identification of protein level and function level showed that the selected BaF3/Tel-IGF1R cell line could correctly express ligand independent tyrosine kinase IGF1R, which laid a good foundation for further drug screening. Drug screening of tyrosine kinase IGF1R cell model. The inhibitory components of tyrosine kinase IGF1R were screened from the natural product library by using the constructed BaF3/Tel-IGF1R cell line as a cell model of high throughput screening. The results of high throughput screening were preliminarily analyzed. The tyrosine kinase IGF1R inhibitory component in the natural product library was confirmed by further validation, which provided guidance for the screening of its specific inhibitors in the future. Furthermore, it can provide lead compounds for the development and research of anti-tumor drugs.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R346

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