靶向IGF-1R单克隆抗体对肺癌的作用机制及增效作用研究

发布时间：2018-06-12 00:24

  本文选题：肺癌 + 胰岛素样生长因子-1(IGF-1)　； 参考：《山东大学》2016年博士论文

【摘要】：第一部分 循环血IGF-1及IGFBP-3水平与肺癌发病风险研究目的：胰岛素样生长因子(IGF)系统在肿瘤形成中发挥重要作用,而肺癌是发病率和死亡率最高的恶性肿瘤之一。我们开展了一项Meta分析探讨循环IGF-1及IGFBP-3水平与肺癌发病风险的关联性。方法：我们就与肺癌患者循坏IGF-1及IGFBP-3水平相关的前瞻性病例对照试验和病例对照试验的研究进行了系统的文献检索。经过严格的纳入和排除标准筛选,共纳入6项巢式病例对照研究(1043例病人及11472例对照)和8项病例对照研究(401例病人及343例正常对照)。我们以多因素校正的OR值及其95%CIs来合并计算循环IGF-1和IGFBP-3水平与肺癌发生率之间的关联性同时,我们计算了标准化均差(SMD)来比较病例组和对照组循环IGF-1和IGFBP-3的差异。结果：对于所有巢式病例对照研究,IGF-1和IGFBP-3水平最高组与最低组比较的合并OR值分别为1.047(95%CI：[0.802,1.367],P=0.736)和0.960(95%CI：[0.591,1.559],P=0.868),结果表明循环血IGF-1、IGFBP3水平与肺癌发生风险不存在统计意义；与合并OR值一致,SMDs分别为-0.079(95%CI：[-0.169,0.011],P=0.086)和-0.097(95%CI：[-0.264,0.071],P=0.258),也不能说明循环血IGF-1、IGFBP3水平与肺癌发生风险之间的关联性。病例对照研究中,循环血IGF-1水平的合并SMDs为0.568(95%CI：[-0.035,1.171],P=0.065),未发现统计学差异； 而值得我们注意的是循环血IGFBP-3的合并SMDs为-0.780(95%CI：[-1.358,-0.201],P=0.008),表明循环血IGFBP-3水平与肺癌发生成负相关。结合巢式病例对照研究和病例对照研究的特点,我们认为巢式病例对照研究的结果代表肺癌患者确诊多年以前血液标本的IGF-1及IGFBP-3水平与一直为患肺癌正常人群之间差异比较,而病理对照研究的结果代表肺癌患者患病状态下血液标本中IGF-1及IGFBP-3水平与正常人血中的差异,我们推断肺癌患者血液IGFBP-3水平在肺癌发生发展过程中有明显的下降过程。结论：病例对照研究结果表明循环IGFBP-3水平与肺癌发生成负相关,且在肺癌的发生发展过程中IGFBP-3水平发生明显下降；这些结果提示IGFBP-3可能成为肺癌的潜在生物标志物。第二部分靶向IGF-1R单克隆抗体在小细胞肺癌中的作用机制及增效作用研究目的：小细胞肺癌是一种具有独特生物学特性的高度恶性肿瘤。靶向治疗抗体成为近来药物研发的热点。本研究拟通过检测小细胞肺癌患者肿瘤组织IGF-R表达情况,并结合患者临床病理资料进行统计学分析,进一步明确IGF-R在小细胞肺癌发生发展中的作用和针对IGF-R治疗的意义。进而我们拟探讨靶向IGF-R单克隆抗体Figitumumab对小细胞肺癌的作用及具体机制；并尝试将MER/ERK抑制剂和二甲双胍分别与Figitumumab联合治疗,观察其增效作用、探索可能的作用机制。期待能够为小细胞肺癌靶向IGF-R单克隆抗体的治疗提供理论基础和联合治疗新思路。方法：我们用免疫组化方法检测了61名小细胞肺癌患者组织标本IGF-1R的表达情况并进行了预后相关性分析。同时,我们用MTT法验证了靶向IGF-1R的单克隆抗体Figitumumab对小细胞肺癌的抗肿瘤效应。采用Western方法检测了IGF-R磷酸化状态及下游PI3K/AKT、MEK/ERK通路的激活情况。并关注了Figitumumab对IGF-R内吞下调的影响。进而,我们尝试了与两种药物的联合治疗以期提高Figitumumab疗效。结果：我们的临床数据显示高表达IGF-1R的小细胞肺癌患者预后较差。进一步实验结果表明Figitumumab对小细胞肺癌细胞系的抑制作用是通过对IGF-1R的受体阻断效应和在不依赖IGF-1R磷酸化及下游PI3K/AKT通路激活的情况下下调IGF-1R实现的。我们发现Figitumumab作用于小细胞肺癌细胞系可激活MEK/ERK信号通路,而通过沉默β-arrestinl可以增强MEK/ERK信号通路激活、沉默β-arrestin2则可减弱该通路。我们发现MEK/ERK抑制剂Figitumumab联合Figitumumab治疗小细胞肺癌可增加疗效,说明抑制ERK通路是发挥增效作用的机制。二甲双胍联合Figitumumab也可增加对小细胞肺癌的抑制作用,我们进一步揭示了二甲双胍的增效作用是通过下调IGF-1R实现的。结论：我们的实验结果验证了靶向IGF-1R单克隆抗体的疗效,提示了MEK/ERK抑制剂和二甲双胍作为联合应用佐剂的应用前景,为后续研究打下基础。第三部分靶向IGF-1R单克隆抗体对非小细胞肺癌的作用机制及增效作用研究目的：胰岛素样生长因子(IGF)信号体系在肿瘤发生发展中发挥重要作用,因此IGF-R成为抗肿瘤治疗中潜在靶点。尽管目前已研发出多重靶向IGF-1R的单克隆抗体药物,验证其抗肿瘤效应和进一步明确其发挥作用的具体分子子机制仍任重道远。本研究拟通过验证靶向IGF-1R的单克隆抗体药物Figitumumab对非小细胞肺癌的作用,并探讨其作用机制。同时,为了探索增加靶向IGF-R治疗疗效的新途径,我们尝试将MEK/ERK抑制剂和二甲双胍分别与Figitumumab联合应用。方法：我们用MTT方法检测Figitumumab对一系列非小细胞肺癌的抑制作用。Western方法检测Figitumumab作用后IGF-1R信号通路的激活情况和IGF-1R内吞下调情况。同时,应用MTT和Western方法检测了MEK/ERK抑制剂和二甲双胍两盒靶向IGF-1R单克隆抗体治疗的增效作用。结果：研究结果证实靶向IGF-1R单克隆抗体Figitumumab对纳入的非小细胞肺癌细胞系均有抑制作用,Figitumumab可阻断IGF-1R磷酸化及下游PI3K/AKT通路的激活。出乎我们意料的是,我们发现Figitumumab可以不依赖IGF-R的激酶活性而激活ERK。进一步研究表明,该ERK的激活是由β-arrestin2参与介导的。同时,我们尝试将MEK/ERK抑制剂与Figitumumab联合应用观察是否有增效作用。我们发现U0126与Figitumumab联合应用可提高对非小细胞肺癌细胞的抑制作用。我们还探讨了二甲双胍的看肿瘤效应并尝试将二甲双胍与Figitumumab联合应用治疗非小细胞肺癌。我们发现二甲双胍可以降低IGF-1R磷酸化及下游PI3K/AKT、MEK/ERK信号转导通路,且可以显著下调IGF-1R表达,我们认为这可能是二甲双胍发挥抗肿瘤作用的机制之一。我们将二甲双胍与Figitumumab联合应用有更显著的抗肿瘤作用,我们认为增加IGF-1R下调可能是该增效作用的机制。结论：本研究验证了靶向IGF-R单克隆抗体对非小细胞肺癌的抑制作用,并探讨了其在阻断信号通路和诱导受体内吞下调方面的作用机制。我们的结果支持二甲双胍可以与抗IGF-1R单克隆抗体联合应用治疗非小细胞肺癌发挥增效作用。第四部分靶向IGF-1R抗体与化疗联合应用治疗肿瘤的协同作用目的：胰岛素样生长因子受体(IGF-1R)是由两个α亚基和两个p亚基组成的细胞膜受体,是IGF-1R信号转导体系的重要成员。IGF-1R体系在细胞增殖、分化、凋亡和表型转化方面发挥重要作用,是导致肿瘤细胞生长失控的原因之一。近年来,许多临床前期实验和临床试验结果表明,将靶向IGF-1R单克隆抗体与化疗药物联合应用可在治疗肿瘤方面有明显的协同增效作用。本研究拟通过整合既往靶向IGF-1R抗体与化疗联合应用的相关临床试验结果,总结其安全性、耐受性、抗肿瘤活性和不良反应等,以期为后续靶向IGF-1R抗体研发和临床试验设计和开展提供理论数据支持。方法：我们对IGF-1R在肿瘤发生发展中发挥的重要作用进行了综述。并整合了针对靶向IGF-1R抗体药物的临床前期实验的结果。进一步,我们系统检索了本研究总结整理了上线和发表的关于靶向IGF-R抗体与化疗联合应用临床试验,提取其中安全性、耐受性、抗肿瘤活性和不良反应等相关数据,进行整合综述。结果：IGF-1R体系在肿瘤发生发展中的举足轻重作用使之成为肿瘤治疗中很有潜力的靶点之一。同时,IGF-R信号转导体系在多种临床有效的抗肿瘤药物的耐药中发挥重要作用。与传统的标准化疗相比,在多重肿瘤中的联合用药均可明显增加患者无进展生存期、客观缓解率和疾病稳定。然而,并非所有联合用药的临床试验都得到了令人满意的效果。临床试验中出现的不良反应多数都是轻微可控的,其中高糖血症是发生率最高的不良反应。结论：一些Ⅰ/Ⅱ临床试验结果表明靶向IGF-1R抗体药物与传统化疗联合应用可发挥协同增效作用,然而,仍有部分临床试验效果不佳。据临床前期实验的令人振奋的抗肿瘤结果而言,我们依然对靶向IGF-1R治疗及其联合治疗有很大信心。这种挑战与机遇并存的现状,也要求我们进一步深入探讨药物作用的具体分子机制、探索针对药物敏感性和耐药性的有效生物学标志物、寻找新的治疗方案。
[Abstract]:The first part of the circulating blood IGF-1 and IGFBP-3 level and the risk of lung cancer: the insulin like growth factor (IGF) system plays an important role in the formation of tumor, and lung cancer is one of the most malignant tumors with the highest incidence and mortality. We have carried out a Meta analysis to explore the level of circulating IGF-1 and IGFBP-3 and the risk of lung cancer Methods: We conducted systematic literature searches on prospective case-control and case-control studies in patients with lung cancer associated with bad IGF-1 and IGFBP-3 levels. Through strict inclusion and exclusion criteria, 6 nested case control studies (1043 patients and 11472 controls) and 8 cases were included. According to the study (401 patients and 343 normal controls), we combined the correlation between the OR and the 95%CIs to combine the computed cycle IGF-1 and IGFBP-3 levels with the incidence of lung cancer. We calculated the normalized difference (SMD) to compare the difference between the IGF-1 and the IGFBP-3 in the case group and the control group. Results: for all nests In case control study, the combined values of IGF-1 and IGFBP-3 in the highest levels of IGF-1 and IGFBP-3 were 1.047 (95%CI:[0.802,1.367], P=0.736) and 0.960 (95%CI:[0.591,1.559], P=0.868). The results showed that the level of circulating blood IGF-1, IGFBP3 was not statistically significant to the risk of lung cancer; [-0.169,0.011], P=0.086) and -0.097 (95%CI:[-0.264,0.071], P=0.258) did not explain the association between circulating blood IGF-1, IGFBP3 level and the risk of lung cancer. In a case-control study, the level of circulating blood IGF-1 combined SMDs was 0.568 (95%CI:[-0.035,1.171], P= 0.065), and no statistical difference was found; but it was worth our attention. The combined SMDs of circulating blood IGFBP-3 is -0.780 (95%CI:[-1.358, -0.201], P=0.008), indicating a negative correlation between the level of circulating blood IGFBP-3 and the occurrence of lung cancer. Combined with the characteristics of nested case control study and case control study, we believe that the results of nested case control study represent the IGF-1 and IGFBP- of the blood specimens of the lung cancer patients before years of diagnosis. The difference between the 3 level and the normal people with lung cancer was compared. The results of the pathological study represented the difference between the levels of IGF-1 and IGFBP-3 in the blood specimens of the patients with lung cancer and the normal human blood. We concluded that the blood IGFBP-3 level in the patients with lung cancer had a significant decline during the development of lung cancer. The results of the control study showed that the level of circulating IGFBP-3 was negatively related to the occurrence of lung cancer, and the level of IGFBP-3 decreased significantly during the development of lung cancer. These results suggest that IGFBP-3 may be a potential biomarker for lung cancer. Second the mechanism and synergism of the target IGF-1R monoclonal antibody in small cell lung cancer Objective: small cell lung cancer is a highly malignant tumor with unique biological characteristics. Targeted therapy antibody has become a hot spot in recent drug research and development. This study is to detect the expression of IGF-R in tumor tissues of small cell lung cancer patients and to make statistical analysis in combination with the clinical and pathological data of patients, to further clarify IGF-R in small and thin sections. The role in the development of cell lung cancer and the significance of IGF-R treatment, and then we intend to explore the role and specific mechanism of targeted IGF-R monoclonal antibody Figitumumab for small cell lung cancer, and try to combine MER/ERK inhibitors and metformin with Figitumumab to observe their synergistic effect and explore possible mechanisms of action. We can provide theoretical basis and new combined therapy for the treatment of IGF-R monoclonal antibody to small cell lung cancer. Methods: we detected the expression of IGF-1R in tissue specimens of 61 small cell lung cancer patients by immunohistochemical method and analyzed the correlation of prognosis. At the same time, we verified the monoclonal anti - IGF-1R target by MTT method. The antitumor effect of body Figitumumab on small cell lung cancer. The Western method was used to detect the phosphorylation of IGF-R and the activation of the downstream PI3K/AKT, MEK/ERK pathway. And the effect of Figitumumab on the down regulation of IGF-R endocytosis was concerned. Furthermore, we tried to combine with two drugs to improve the curative effect of Figitumumab. The clinical data showed that the prognosis of small cell lung cancer patients with high expression of IGF-1R was poor. Further experimental results showed that the inhibitory effect of Figitumumab on the cell line of small cell lung cancer was achieved through the blocking effect on IGF-1R receptor and the downregulation of IGF-1R without IGF-1R phosphorylation and the activation of the downstream PI3K/AKT pathway. We found that Figit The effect of umumab on the cell line of small cell lung cancer activates the MEK/ERK signaling pathway, and the silence of beta -arrestinl can enhance the activation of the MEK/ERK signaling pathway. The silence of beta -arrestin2 can weaken the pathway. We found that the MEK/ERK inhibitor Figitumumab combined with Figitumumab in the treatment of small cell lung cancer can increase the efficacy, indicating that the suppression of the ERK pathway is increased. The synergistic mechanism. Metformin combined with Figitumumab can also increase the inhibitory effect on small cell lung cancer. We further revealed that the synergistic effect of metformin is achieved by down regulation of IGF-1R. Conclusion: our experimental results verify the efficacy of the target IGF-1R monoclonal antibody, suggesting that the MEK/ERK inhibitor and metformin are used as a result. The application prospect of combined application of adjuvant has laid a foundation for follow-up research. Third the mechanism and synergism of target IGF-1R monoclonal antibody against non-small cell lung cancer. Objective: the insulin like growth factor (IGF) signal system plays an important role in the development of tumor. Therefore, IGF-R has become a potential target in anti-tumor therapy. Although the monoclonal antibody against IGF-1R has been developed, it is still a long way to go to verify its anti-tumor effect and further clarify its specific molecular mechanism. This study is to verify the effect of monoclonal antibody drug Figitumumab targeting IGF-1R on non small cell lung cancer and to explore its mechanism. In order to explore new ways to increase the therapeutic effect of targeted IGF-R therapy, we try to combine MEK/ERK inhibitors and metformin with Figitumumab. Methods: we use MTT to detect the inhibitory effect of Figitumumab on a series of non small cell lung cancers by.Western method to detect the activation of IGF-1R signaling pathway and I after Figitumumab action. Down regulation of GF-1R endocytosis. At the same time, MTT and Western methods were used to detect the synergistic effect of MEK/ERK inhibitors and metformin two boxes targeting IGF-1R monoclonal antibody. Results: the results showed that the targeted IGF-1R monoclonal antibody Figitumumab had inhibitory effect on the included non-small cell lung cancer cell lines, and Figitumumab could block IGF-1. R phosphorylation and downstream PI3K/AKT pathway activation. To our expectation, we found that Figitumumab can activate ERK. without dependence on IGF-R kinase activity and further studies show that the activation of the ERK is mediated by beta -arrestin2. Meanwhile, we try to observe whether there is a synergistic effect by combining MEK/ERK inhibitors with Figitumumab. We found that the combination of U0126 and Figitumumab can improve the inhibition of non-small cell lung cancer cells. We also explored the tumor effect of metformin and the combination of metformin and Figitumumab in the treatment of non small cell lung cancer. We found that metformin can lower IGF-1R phosphorylation and downstream PI3K/AKT, MEK/ERK The signal transduction pathway, which can significantly downregulate the expression of IGF-1R, may be one of the mechanisms that metformin plays an anti-tumor role. We have combined the combination of metformin and Figitumumab to have a more significant antitumor effect. We think that the increase of IGF-1R may be the mechanism of this effect. Conclusion: This study verified the target. The inhibitory effect of IGF-R monoclonal antibody on non-small cell lung cancer and the mechanism of its role in blocking signal pathways and inducing endocytosis in vivo. Our results support the synergistic effect of metformin in combination with anti IGF-1R monoclonal antibodies in the treatment of non-small cell lung cancer. Fourth part of the target IGF-1R antibody and The synergistic effect of combined chemotherapy in the treatment of tumors: the insulin like growth factor receptor (IGF-1R) is a cell membrane receptor composed of two subunits and two p subunits. It is an important member of the IGF-1R signal transduction system,.IGF-1R system plays an important role in cell proliferation, differentiation, apoptosis and surface transformation, and is the cause of tumor cells. One of the reasons for uncontrollable growth. In recent years, many preclinical and clinical trials have shown that the combination of targeted IGF-1R monoclonal antibodies and chemotherapeutic drugs can synergistically synergistically in the treatment of tumors. This study is intended to integrate the relevant clinical trial results of the combination of previous targeted IGF-1R antibodies and chemotherapy. To summarize its safety, tolerance, antitumor activity and adverse reactions, in order to provide theoretical data support for the design and development of subsequent target IGF-1R antibody development and clinical trials. Methods: We reviewed the important role of IGF-1R in the development of tumor and integrated the clinical prophase of targeted IGF-1R antibody drugs. The results of the experiment. Further, we systematically retrieved this study to summarize the online and published clinical trials of the combined use of targeted IGF-R antibody and chemotherapy to extract the data related to safety, tolerance, antitumor activity and adverse reactions. Results: the IGF-1R system was used in the development of cancer. The light and heavy effects make it one of the potential targets in cancer treatment. At the same time, the IGF-R signal transduction system plays an important role in the resistance of a variety of clinically effective antitumor drugs. Compared with traditional standard chemotherapy, combined use in multiple tumors can significantly increase the patient's progression free survival, objective remission rate and disease. It is stable. However, not all clinical trials of combined use of drugs have been satisfactory. Most of the adverse reactions in clinical trials are slightly controlled, and hyperglycemia is the highest incidence of adverse reactions. Conclusions: some I / II clinical trials show that targeted IGF-1R antibody drugs are combined with traditional chemotherapy. However, some of the clinical trials still have a poor effect. According to the exciting antitumor results of pre clinical trials, we still have great confidence in the targeting of IGF-1R therapy and its combined treatment. The present situation of the challenge and opportunity also requires us to further explore the specific effects of the drug. Molecular mechanisms to explore effective biomarkers for drug sensitivity and drug resistance, and search for new therapeutic options.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2
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