本文選題：肺癌 + 胰島素樣生長因子-1(IGF-1)��； 參考：《山東大學》2016年博士論文

【摘要】：第一部分 循環(huán)血IGF-1及IGFBP-3水平與肺癌發(fā)病風險研究目的：胰島素樣生長因子(IGF)系統(tǒng)在腫瘤形成中發(fā)揮重要作用,而肺癌是發(fā)病率和死亡率最高的惡性腫瘤之一。我們開展了一項Meta分析探討循環(huán)IGF-1及IGFBP-3水平與肺癌發(fā)病風險的關聯(lián)性。方法：我們就與肺癌患者循壞IGF-1及IGFBP-3水平相關的前瞻性病例對照試驗和病例對照試驗的研究進行了系統(tǒng)的文獻檢索。經(jīng)過嚴格的納入和排除標準篩選,共納入6項巢式病例對照研究(1043例病人及11472例對照)和8項病例對照研究(401例病人及343例正常對照)。我們以多因素校正的OR值及其95%CIs來合并計算循環(huán)IGF-1和IGFBP-3水平與肺癌發(fā)生率之間的關聯(lián)性同時,我們計算了標準化均差(SMD)來比較病例組和對照組循環(huán)IGF-1和IGFBP-3的差異。結果：對于所有巢式病例對照研究,IGF-1和IGFBP-3水平最高組與最低組比較的合并OR值分別為1.047(95%CI：[0.802,1.367],P=0.736)和0.960(95%CI：[0.591,1.559],P=0.868),結果表明循環(huán)血IGF-1、IGFBP3水平與肺癌發(fā)生風險不存在統(tǒng)計意義；與合并OR值一致,SMDs分別為-0.079(95%CI：[-0.169,0.011],P=0.086)和-0.097(95%CI：[-0.264,0.071],P=0.258),也不能說明循環(huán)血IGF-1、IGFBP3水平與肺癌發(fā)生風險之間的關聯(lián)性。病例對照研究中,循環(huán)血IGF-1水平的合并SMDs為0.568(95%CI：[-0.035,1.171],P=0.065),未發(fā)現(xiàn)統(tǒng)計學差異； 而值得我們注意的是循環(huán)血IGFBP-3的合并SMDs為-0.780(95%CI：[-1.358,-0.201],P=0.008),表明循環(huán)血IGFBP-3水平與肺癌發(fā)生成負相關。結合巢式病例對照研究和病例對照研究的特點,我們認為巢式病例對照研究的結果代表肺癌患者確診多年以前血液標本的IGF-1及IGFBP-3水平與一直為患肺癌正常人群之間差異比較,而病理對照研究的結果代表肺癌患者患病狀態(tài)下血液標本中IGF-1及IGFBP-3水平與正常人血中的差異,我們推斷肺癌患者血液IGFBP-3水平在肺癌發(fā)生發(fā)展過程中有明顯的下降過程。結論：病例對照研究結果表明循環(huán)IGFBP-3水平與肺癌發(fā)生成負相關,且在肺癌的發(fā)生發(fā)展過程中IGFBP-3水平發(fā)生明顯下降；這些結果提示IGFBP-3可能成為肺癌的潛在生物標志物。第二部分靶向IGF-1R單克隆抗體在小細胞肺癌中的作用機制及增效作用研究目的：小細胞肺癌是一種具有獨特生物學特性的高度惡性腫瘤。靶向治療抗體成為近來藥物研發(fā)的熱點。本研究擬通過檢測小細胞肺癌患者腫瘤組織IGF-R表達情況,并結合患者臨床病理資料進行統(tǒng)計學分析,進一步明確IGF-R在小細胞肺癌發(fā)生發(fā)展中的作用和針對IGF-R治療的意義。進而我們擬探討靶向IGF-R單克隆抗體Figitumumab對小細胞肺癌的作用及具體機制；并嘗試將MER/ERK抑制劑和二甲雙胍分別與Figitumumab聯(lián)合治療,觀察其增效作用、探索可能的作用機制。期待能夠為小細胞肺癌靶向IGF-R單克隆抗體的治療提供理論基礎和聯(lián)合治療新思路。方法：我們用免疫組化方法檢測了61名小細胞肺癌患者組織標本IGF-1R的表達情況并進行了預后相關性分析。同時,我們用MTT法驗證了靶向IGF-1R的單克隆抗體Figitumumab對小細胞肺癌的抗腫瘤效應。采用Western方法檢測了IGF-R磷酸化狀態(tài)及下游PI3K/AKT、MEK/ERK通路的激活情況。并關注了Figitumumab對IGF-R內(nèi)吞下調(diào)的影響。進而,我們嘗試了與兩種藥物的聯(lián)合治療以期提高Figitumumab療效。結果：我們的臨床數(shù)據(jù)顯示高表達IGF-1R的小細胞肺癌患者預后較差。進一步實驗結果表明Figitumumab對小細胞肺癌細胞系的抑制作用是通過對IGF-1R的受體阻斷效應和在不依賴IGF-1R磷酸化及下游PI3K/AKT通路激活的情況下下調(diào)IGF-1R實現(xiàn)的。我們發(fā)現(xiàn)Figitumumab作用于小細胞肺癌細胞系可激活MEK/ERK信號通路,而通過沉默β-arrestinl可以增強MEK/ERK信號通路激活、沉默β-arrestin2則可減弱該通路。我們發(fā)現(xiàn)MEK/ERK抑制劑Figitumumab聯(lián)合Figitumumab治療小細胞肺癌可增加療效,說明抑制ERK通路是發(fā)揮增效作用的機制。二甲雙胍聯(lián)合Figitumumab也可增加對小細胞肺癌的抑制作用,我們進一步揭示了二甲雙胍的增效作用是通過下調(diào)IGF-1R實現(xiàn)的。結論：我們的實驗結果驗證了靶向IGF-1R單克隆抗體的療效,提示了MEK/ERK抑制劑和二甲雙胍作為聯(lián)合應用佐劑的應用前景,為后續(xù)研究打下基礎。第三部分靶向IGF-1R單克隆抗體對非小細胞肺癌的作用機制及增效作用研究目的：胰島素樣生長因子(IGF)信號體系在腫瘤發(fā)生發(fā)展中發(fā)揮重要作用,因此IGF-R成為抗腫瘤治療中潛在靶點。盡管目前已研發(fā)出多重靶向IGF-1R的單克隆抗體藥物,驗證其抗腫瘤效應和進一步明確其發(fā)揮作用的具體分子子機制仍任重道遠。本研究擬通過驗證靶向IGF-1R的單克隆抗體藥物Figitumumab對非小細胞肺癌的作用,并探討其作用機制。同時,為了探索增加靶向IGF-R治療療效的新途徑,我們嘗試將MEK/ERK抑制劑和二甲雙胍分別與Figitumumab聯(lián)合應用。方法：我們用MTT方法檢測Figitumumab對一系列非小細胞肺癌的抑制作用。Western方法檢測Figitumumab作用后IGF-1R信號通路的激活情況和IGF-1R內(nèi)吞下調(diào)情況。同時,應用MTT和Western方法檢測了MEK/ERK抑制劑和二甲雙胍兩盒靶向IGF-1R單克隆抗體治療的增效作用。結果：研究結果證實靶向IGF-1R單克隆抗體Figitumumab對納入的非小細胞肺癌細胞系均有抑制作用,Figitumumab可阻斷IGF-1R磷酸化及下游PI3K/AKT通路的激活。出乎我們意料的是,我們發(fā)現(xiàn)Figitumumab可以不依賴IGF-R的激酶活性而激活ERK。進一步研究表明,該ERK的激活是由β-arrestin2參與介導的。同時,我們嘗試將MEK/ERK抑制劑與Figitumumab聯(lián)合應用觀察是否有增效作用。我們發(fā)現(xiàn)U0126與Figitumumab聯(lián)合應用可提高對非小細胞肺癌細胞的抑制作用。我們還探討了二甲雙胍的看腫瘤效應并嘗試將二甲雙胍與Figitumumab聯(lián)合應用治療非小細胞肺癌。我們發(fā)現(xiàn)二甲雙胍可以降低IGF-1R磷酸化及下游PI3K/AKT、MEK/ERK信號轉導通路,且可以顯著下調(diào)IGF-1R表達,我們認為這可能是二甲雙胍發(fā)揮抗腫瘤作用的機制之一。我們將二甲雙胍與Figitumumab聯(lián)合應用有更顯著的抗腫瘤作用,我們認為增加IGF-1R下調(diào)可能是該增效作用的機制。結論：本研究驗證了靶向IGF-R單克隆抗體對非小細胞肺癌的抑制作用,并探討了其在阻斷信號通路和誘導受體內(nèi)吞下調(diào)方面的作用機制。我們的結果支持二甲雙胍可以與抗IGF-1R單克隆抗體聯(lián)合應用治療非小細胞肺癌發(fā)揮增效作用。第四部分靶向IGF-1R抗體與化療聯(lián)合應用治療腫瘤的協(xié)同作用目的：胰島素樣生長因子受體(IGF-1R)是由兩個α亞基和兩個p亞基組成的細胞膜受體,是IGF-1R信號轉導體系的重要成員。IGF-1R體系在細胞增殖、分化、凋亡和表型轉化方面發(fā)揮重要作用,是導致腫瘤細胞生長失控的原因之一。近年來,許多臨床前期實驗和臨床試驗結果表明,將靶向IGF-1R單克隆抗體與化療藥物聯(lián)合應用可在治療腫瘤方面有明顯的協(xié)同增效作用。本研究擬通過整合既往靶向IGF-1R抗體與化療聯(lián)合應用的相關臨床試驗結果,總結其安全性、耐受性、抗腫瘤活性和不良反應等,以期為后續(xù)靶向IGF-1R抗體研發(fā)和臨床試驗設計和開展提供理論數(shù)據(jù)支持。方法：我們對IGF-1R在腫瘤發(fā)生發(fā)展中發(fā)揮的重要作用進行了綜述。并整合了針對靶向IGF-1R抗體藥物的臨床前期實驗的結果。進一步,我們系統(tǒng)檢索了本研究總結整理了上線和發(fā)表的關于靶向IGF-R抗體與化療聯(lián)合應用臨床試驗,提取其中安全性、耐受性、抗腫瘤活性和不良反應等相關數(shù)據(jù),進行整合綜述。結果：IGF-1R體系在腫瘤發(fā)生發(fā)展中的舉足輕重作用使之成為腫瘤治療中很有潛力的靶點之一。同時,IGF-R信號轉導體系在多種臨床有效的抗腫瘤藥物的耐藥中發(fā)揮重要作用。與傳統(tǒng)的標準化療相比,在多重腫瘤中的聯(lián)合用藥均可明顯增加患者無進展生存期、客觀緩解率和疾病穩(wěn)定。然而,并非所有聯(lián)合用藥的臨床試驗都得到了令人滿意的效果。臨床試驗中出現(xiàn)的不良反應多數(shù)都是輕微可控的,其中高糖血癥是發(fā)生率最高的不良反應。結論：一些Ⅰ/Ⅱ臨床試驗結果表明靶向IGF-1R抗體藥物與傳統(tǒng)化療聯(lián)合應用可發(fā)揮協(xié)同增效作用,然而,仍有部分臨床試驗效果不佳。據(jù)臨床前期實驗的令人振奮的抗腫瘤結果而言,我們依然對靶向IGF-1R治療及其聯(lián)合治療有很大信心。這種挑戰(zhàn)與機遇并存的現(xiàn)狀,也要求我們進一步深入探討藥物作用的具體分子機制、探索針對藥物敏感性和耐藥性的有效生物學標志物、尋找新的治療方案。
[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.
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R734.2
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本文編號：2007359