Notch1、IGF-1在非小细胞肺癌EGFR不同基因状态下的表达及临床意义

发布时间：2018-04-22 05:37

本文选题：Notch1 + IGF-1　；参考：《济南大学》2017年硕士论文

【摘要】：研究背景随着医疗水平的不断提升,特别是在分子生物学技术的研究方面,人们对非小细胞肺癌的发病机制有了更深层次的了解,而在非小细胞肺癌的药物治疗研究上,靶点表皮生长因子受体(epidermal growth factor receptor,EGFR)的发现具有里程碑意义,尤其是对于那些晚期肺癌患者,靶向治疗给他们带来了福音,同时也使得非小细胞肺癌的药物治疗进入了一个全新时代。EGFR是Erb B受体家族的其中一员,该受体家族共有四种受体,分别称为EGFR(Erb B-1或HER-1)、Erb B-2(HER-2)、Erb B-3(HER-3)和Erb B-4(HER-4)。EGFR发生基因突变或表达过度会影响肿瘤细胞的发生发展。随之,EGFR抑制剂便应运而生,它针对EGFR为靶点发挥治疗作用,因此特异性高,而且高效;EGFR抑制剂主要分为两类:即小分子酪氨酸激酶抑制剂(tyrosine kinases inhibitor,TKI)和单克隆抗体(Mab),前者的药理机制是切割EGFR胞内段的酪氨酸激酶结构域与ATP相连接,使磷酸化失败,并进一步阻扰了下游信号的激活,主要有两条:MAPK和PI3K/Akt;后者的药理作用是使EGFR与相应配体结合失败,进而EGFR二聚化受阻,从而切断该条致癌信号通路。EGFR受体与相应的配体结合可活化EGFR信号通路,促进肿瘤细胞的增生和正常细胞的转化,抑制细胞程序化凋亡,加快肿瘤组织的血管生成,致使肿瘤细胞浸润、转移,最终导致肿瘤发生、发展。EGFR-TKI可抑制肿瘤浸润和转移,同时可提高肿瘤组织放射敏感性,治疗效果显著。但临床上发现,EGFR-TKI治疗数月后即产生耐药,因此研究其中耐药机制成为热点。既往研究表明肺癌的发生与Notch1信号通路的激活密切相关,而EGFR-TKI耐药是否也Notch1参与有目前尚无研究明确指出,但研究发现Notch-1是细胞内众多信号通路的交汇点,因此,EGFR信号通路下游的转导通路中很可能与Notch1也存在交叉作用。国外有研究表明Notch-1的促肿瘤生存的功能是通过激活胰岛素样生长因子-1受体(insulin-like growth factors,IGF-1R)来发挥作用的,但目前对于两者的表达相关性分析较少,本研究致力于此,并探讨Notch1在非小细胞肺癌靶向药物耐药机制,希望能为EGFR-TKI耐药机制研究寻找出新的突破点,为靶向药物治疗提供新的思路。目的运用免疫组化法检测非小细胞肺癌中Notch1与IGF-1表达情况,分析两者在EGFR野生组、EGFR突变组以及EGFR耐药组中表达的相关性及临床意义,并进一步探讨EGFR-TKI耐药机制,为非小细胞肺癌靶向药物耐药研究提供新途径。方法回顾性分析85例NSCLC患者基本病历资料,按照EGFR基因状态的不同划分为三组,即野生组、突变组和耐药组,采用免疫组化SP法分别对38例EGFR野生组、28例EGFR突变组,19例耐药组进行Notch1、IGF-1的蛋白表达检测。结果野生组中非小细胞肺癌Notch1、IGF-1阳性表达率分别为26.34%、21.05%,突变组中非小细胞肺癌Notch1、IGF-1阳性表达率分别为39.29%、32.14%,而耐药组中非小细胞肺癌Notch1、IGF-1阳性表达率则分别为47.39%、42.11%。Notch1在耐药组中表达明显高于野生组和突变组,其差异存在统计学意义,F=42.003(P0.001);IGF-1在耐药组中的表达明显高于野生组和突变组,其差异存在统计学意义,F=38.179(P0.001)。随着非小细胞肺癌病理分级的升高和分化程度降低,Notch-1和IGF-1蛋白在三组中的阳性表达率均增高(P0.05);野生组、突变组、耐药组各组中Notch1与IGF-1的相关性分析,结果显示野生组与突变组中两者的表达无相关性(野生组:r=0.278 P=0.091,突变组:r=0.229 P=0.241),差异无统计学意义;但在耐药组中Notch1与IGF-1呈正相关(r=0.685 P=0.001),差异有统计学意义。结论本研究显示:EGFR耐药组中Notch1的表达明显高于其他两组且差异存在统计学意义(P0.05);EGFR耐药组中IGF-1的表达明显高于其他两组且差异存在统计学意义(P0.05);EGFR耐药组Notch1、IGF-1表达具有相关性,在其他两组中无相关性;结合国内外相关研究推测Notch1导致EGFR-TKI耐药的原因可能是先激活IGF-1,然后进一步激活EGFR下游的Ras/MAPK和P13K/AKT信号通路来完成的,提示Notch1信号高表达可能为EGFR-TKIs耐药的重要因素。
[Abstract]:Background with the continuous improvement of medical level, especially in the study of molecular biology, people have a deeper understanding of the pathogenesis of non-small cell lung cancer, and the discovery of epidermal growth factor receptor (EGFR) in the drug therapy of non small cell lung cancer has been found. The milestone, especially for patients with advanced lung cancer, has brought them the gospel, and it also makes the drug treatment of non small cell lung cancer enter a new era of.EGFR is one of the Erb B receptor family. There are four receptors in the family, called EGFR (Erb B-1 or HER-1), Erb B-2 (HER-2), Erb B-3 (HER-3) and Erb B-4 (HER-4).EGFR gene mutation or overexpression may affect the development of tumor cells. Accordingly, EGFR inhibitors have emerged as the times require. It is targeted to EGFR as a target and is highly specific and efficient; EGFR inhibitors are mainly divided into two categories: small molecule tyrosine kinase inhibitors (tyrosine kinases inhib) Itor, TKI) and monoclonal antibodies (Mab). The pharmacological mechanism of the former is to connect the tyrosine kinase domain of the EGFR intracellular segment to ATP, which causes the failure of phosphorylation and further obstruct the activation of the downstream signals, mainly two: MAPK and PI3K/Akt; the latter has a pharmacological effect that causes the failure of EGFR to be combined with the corresponding ligands, and thus EGFR dimerization is hindered, The combination of the.EGFR receptor and the corresponding ligand can activate the EGFR signal pathway, promote the proliferation of tumor cells and the transformation of normal cells, inhibit the programmed apoptosis of the cells, accelerate the angiogenesis of the tumor tissue, cause the tumor cells to infiltrate, transfer, and eventually lead to the tumorigenesis, and the development of.EGFR-TKI can inhibit the tumor. Infiltration and metastasis can also improve the radiosensitivity of tumor tissue, and the therapeutic effect is significant. However, it is found that drug resistance occurs after a few months of EGFR-TKI treatment. Therefore, the mechanism of drug resistance has become a hot spot. Previous studies have shown that the occurrence of lung cancer is related to the activation of Notch1 signaling pathway, and whether the drug resistance of EGFR-TKI is also involved in the presence of Notch1. No research has been made before, but the study found that Notch-1 is the intersection of numerous signaling pathways in the cells. Therefore, the downstream transduction pathway in the EGFR signaling pathway is likely to be intersecting with Notch1. Foreign studies have shown that the function of Notch-1 to promote tumor survival is through the stimulated insulin like growth factor -1 receptor (insulin-like GRO). Wth factors, IGF-1R) to play a role, but at present, there is little correlation analysis of the expression of the two. This study is devoted to this, and explores the mechanism of drug resistance of Notch1 in non-small cell lung cancer, hoping to find new breakthrough points for the study of EGFR-TKI resistance mechanism and to provide new ideas for targeting drug therapy. The expression of Notch1 and IGF-1 in non small cell lung cancer was detected by chemical method. The correlation and clinical significance of both in EGFR wild group, EGFR mutation group and EGFR drug resistance group were analyzed, and the mechanism of EGFR-TKI resistance was further explored to provide a new way for the study of drug resistance in non-small cell lung cancer. Methods 85 cases of NSCLC patients were analyzed retrospectively. The data of the medical records were divided into three groups according to the different EGFR gene state, that is, the wild group, the mutant group and the drug resistance group. The immunohistochemical SP method was used to detect the Notch1 and IGF-1 expression of Notch1 and IGF-1 in 38 cases of EGFR wild group, 28 case of EGFR mutation group and 19 cases of drug-resistant group. The positive expression rate of non small cell lung cancer in the wild group was 26.34, respectively. The positive rates of Notch1 and IGF-1 in non small cell lung cancer in the mutant group were 39.29%, 32.14%, respectively, while the Notch1 and IGF-1 positive rates of non small cell lung cancer in the drug resistant group were 47.39%, and 42.11%.Notch1 was significantly higher in the drug resistant group than in the wild group and the mutant group. The difference was statistically significant, F=42.003 (P0.001), and IGF-1 in drug resistance. The expression in the group was significantly higher than that in the wild group and the mutant group. The difference was statistically significant, F=38.179 (P0.001). The positive expression rate of Notch-1 and IGF-1 protein in the three groups increased with the increase of pathological grade and the degree of differentiation of non small cell lung cancer (P0.05), and the correlation between Notch1 and IGF-1 in the wild group, the mutant group and the drug resistant group. The results showed that there was no correlation between the wild group and the mutant group (wild group: r=0.278 P=0.091, mutation group: r=0.229 P=0.241), and there was no significant difference in the difference between Notch1 and IGF-1 (r=0.685 P=0.001) in the drug resistant group (r=0.685 P=0.001), and the difference was statistically significant. Conclusion this study showed that the expression of Notch1 in the EGFR resistant group was significantly higher than that in the drug resistant group. The difference between the other two groups was statistically significant (P0.05), and the expression of IGF-1 in the EGFR resistant group was significantly higher than that of the other two groups, and the difference was statistically significant (P0.05). The EGFR drug resistance group was Notch1, IGF-1 expression was related, and there was no correlation in the other two groups. IGF-1 is activated first and then further activates the Ras/MAPK and P13K/AKT signaling pathways downstream of the EGFR, suggesting that the high expression of Notch1 signals may be an important factor in the resistance of EGFR-TKIs.

【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2

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