本文選題：IGF1R激酶受體　切入點(diǎn)：BaF3細(xì)胞　出處：《華東師范大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：胰島素樣生長(zhǎng)因子1受體(insulin-like growth factor 1 receptor, IGF1R)是酪氨酸蛋白激酶家族的成員,在各種類型細(xì)胞上均有表達(dá)。IGF1R具有潛在的有絲分裂原作用,可促進(jìn)細(xì)胞的增殖、調(diào)節(jié)細(xì)胞的惡性轉(zhuǎn)化、保護(hù)腫瘤細(xì)胞免受凋亡等生物功能,其信號(hào)轉(zhuǎn)導(dǎo)途徑與腫瘤的發(fā)生密切相關(guān),因此IGF1R被認(rèn)為是非常具有潛力的抗腫瘤藥物的靶點(diǎn)。 本論文構(gòu)建酪氨酸激酶IGF1R組成型高表達(dá)的細(xì)胞模型,并以該細(xì)胞模型對(duì)天然產(chǎn)物化合物庫(kù)進(jìn)行高通量藥物篩選,篩選出對(duì)酪氨酸激酶IGF1R具有特異性抑制作用的組分。該結(jié)果進(jìn)一步驗(yàn)證該IGF1R酪氨酸激酶細(xì)胞模型的穩(wěn)定性、可靠性及其于藥物篩選的應(yīng)用。 1 IGF1R酪氨酸激酶細(xì)胞模型構(gòu)建 以實(shí)驗(yàn)室已構(gòu)建的哺乳動(dòng)物細(xì)胞表達(dá)載體pcDNA3.1(+)-Tel為基礎(chǔ),將IGF1R基因與之連接構(gòu)建重組質(zhì)粒。將重組質(zhì)粒轉(zhuǎn)入IL3依賴性的小鼠原B細(xì)胞(BaF3細(xì)胞)并在G418及IL3非依賴性的條件下篩選出能夠組成型高表達(dá)IGF1R激酶活性的細(xì)胞株BaF3/Tel-IGF1R.通過(guò)分別從分子水平,蛋白水平及功能水平的鑒定,說(shuō)明篩選所得到BaF3/Tel-IGF1R細(xì)胞株能夠正確的表達(dá)配體非依賴性的具有組成型活性的酪氨酸激酶IGF1R,為后續(xù)藥物篩選的應(yīng)用打下良好的基礎(chǔ)。 2酪氨酸激酶IGF1R細(xì)胞模型的藥物篩選應(yīng)用 以構(gòu)建的BaF3/Tel-IGF1R細(xì)胞株為高通量篩選的細(xì)胞模型,從天然產(chǎn)物庫(kù)中篩選酪氨酸激酶IGF1R的抑制組分。對(duì)于高通量篩選的結(jié)果進(jìn)行初步分析,并確定有活性的組分進(jìn)一步驗(yàn)證。通過(guò)驗(yàn)證確定天然產(chǎn)物庫(kù)中酪氨酸激酶IGF1R抑制組分,為以后其特異性抑制劑的篩選提供指導(dǎo)意義,進(jìn)而為抗腫瘤藥物的開發(fā)與研究提供先導(dǎo)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.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R346

【引證文獻(xiàn)】

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1 張哲;正電荷修飾的非標(biāo)記核酸熒光探針的研究及其在中藥抗腫瘤研究中的應(yīng)用[D];吉林大學(xué);2012年

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本文編號(hào)：1637351