EGFR和IDO在乳腺癌中共表达的实验研究

发布时间：2018-05-07 08:45

本文选题：EGFR + IDO　；参考：《吉林大学》2015年博士论文

【摘要】：乳腺癌是女性最常见的恶性肿瘤，近年来，全世界乳腺癌发病率不断增高，在我国乳腺癌也呈现不断增高的趋势。尤其值得关注的是乳腺癌目前已成为导致女性死亡的重要因素之一。尽管乳腺癌的诊断与治疗已经取得了很大进展，但是如何有效地控制乳腺癌的进展，延长乳腺癌患者的生存期以及防止或减少乳腺癌的复发，降低乳腺癌的死亡率仍是目前乳腺癌治疗的难题。目前，生物靶向治疗已经是继传统的手术治疗、放疗及化疗后的常规治疗手段之一。包括人源化单抗及小分子靶向药物已经在包括乳腺癌在内的多种癌症治疗当中广泛应用，并且证实对包括乳腺癌在内的多种癌症均有一定的疗效。但是单一靶点治疗仍存在肿瘤易发生变异并产生耐药性，从而导致肿瘤治疗失效的现象发生。如何采用两种或多种靶向药物联合治疗对于提高包括乳腺癌等多种肿瘤的治疗效果具有潜在的应用前景。因此，寻找可联合应用于肿瘤生物治疗的靶点是联合应用靶向药物治疗乳腺癌等多种肿瘤的关键因素之一。以往的研究表明，EGFR在多种肿瘤的发生、发展中具有重要作用，尤其是对于头颈癌、肺癌等多种上皮组织源性的恶性肿瘤。EGFR的过表达对于上述恶性肿瘤的发生、发展、预后转归等均具有重要作用，针对EGFR靶向治疗药物如爱必妥、泰欣生以及小分子靶向药物易瑞沙等的临床应用均有效地抑制了上皮源性的恶性肿瘤组织的生长与转移。在乳腺癌的发生发展过程中，EGFR也同样发挥了重要作用。研究证实，存在于乳腺癌细胞表面EGFR的活化，导致胞内酪氨酸激酶区的活化。另外，EGFR在组织中的过度表达可造成多条信号传导通路的持续性激活，促进蛋白质合成，抑制细胞的凋亡，加速细胞的分裂和增殖，从而促进乳腺癌的发生、发展。因此，EGFR对于乳腺癌的发生、发展及预后转归均起着重要作用，是乳腺癌治疗的靶点之一。吲哚胺2，3-双加氧酶（indoleamine2，3-dioxygenase，IDO）在人及动物代谢中的作用越来越引起研究者的广泛重视，并且广泛分布于人和其他哺乳动物除肝脏以外的组织中， IDO通过降解色氨酸，使局部造成色氨酸缺乏进而T细胞增殖受到抑制。另外，，IDO的降解产物犬尿素及其衍生物也参与免疫调节过程可诱导T细胞凋亡。研究显示，IDO的高表达在包括乳腺癌在内的多种肿瘤中与肿瘤发生免疫逃逸密切相关，因此，IDO是包括乳腺癌在内的多种肿瘤的有研究前景的靶点之一。许多研究报道显示，利用IDO的抑制剂通过抑制IDO发挥作用，使T细胞的增殖恢复到正常状态，从而对抑制肿瘤的浸润转移具有一定的治疗作用。目前，肿瘤生物靶向治疗也逐渐走向深入，从单一靶点的治疗到多靶点以及多种手段联合治疗,并且实践证实,多靶点联合治疗具有减少药物用量，减少肿瘤耐药性发生等诸多优势。因此，多靶点联合肿瘤治疗已经成为肿瘤治疗未来发展的趋势之一。尽管EGFR和IDO在乳腺癌中的研究都分别有报道，但目前尚未见到EGFR和IDO二者抑制剂联合应用干预乳腺癌的发生发展、预后转归进程的相关研究。本文拟通过检测乳腺癌中EGFR与IDO共表达的情况，从而联合抑制EGFR与IDO，为探讨多靶点联合抑制乳腺癌的发展及预后提供新的治疗策略。首先检测乳腺癌患者中EGFR与IDO共表达的分布情况，为实施联合抑制EGFR与IDO双靶点，从而联合抑制乳腺癌的发展预后奠定基础。通过对110例乳腺癌患者标本制成石蜡切片，用免疫组化方法检测，结果证实乳腺癌组织中确实存在EGFR和IDO共表达情况，并且二者共表达情况在乳腺癌患者中占有一定的比例。总共表达率为29.1%，同步共表达19.1%，差异共表达10%。但二者的超微结构表达区域是有差异的。对于IDO，主要表达在恶性导管细胞的细胞浆，在临近的基底细胞大多检测不到。对于EGFR主要表达在上皮细胞和基底细胞的细胞膜上。IDO和EGFR的同步共表达程度由低到高。同时，IDO和EGFR也存在差异共表达，如IDO低表达时，EGFR中度或高表达。而EGFR低表达时，IDO中度或高表达。为进一步通过试验研究证实上述实验结果，我们对8种乳腺癌细胞系进行了检测，利用免疫细胞化学，RT-PCR及Westen Blot的方法相互验证。结果表明，乳腺癌细胞系中也同样存在IDO和EGFR共表达情况。上述两方面试验结果证实开展以EGFR和IDO联合治疗乳腺癌的前提条件是成立的。在此基础上，我们通过体外细胞试验采用艾比妥、易瑞沙、1-甲基色氨酸联合用药，初步证实联合抑制EGFR和IDO的表达能够有效地抑制乳腺癌细胞的生长与侵袭浸润及转移，这为我们今后开展体内试验研究奠定了基础。结论： 1、我们的研究首次证实乳腺癌患者中存在EGFR和IDO的共表达情况，并且存在差异共表达和同步共表达两种情况。 2、我们的研究首次证实乳腺癌细胞系中也同样存在EGFR和IDO的共表达情况。 3、我们的体外试验证实通过对EGFR和IDO联合抑制，能有效地抑制乳腺癌细胞的生长、浸润与转移，并且明显优于单独抑制情况。表明二者联合抑制存在协同效应。
[Abstract]:Breast cancer is the most common malignant tumor in women. In recent years, the incidence of breast cancer in the world is increasing, and the trend of breast cancer is also increasing in China. It is particularly noteworthy that breast cancer has become one of the important factors that lead to the death of women. Although the diagnosis and treatment of breast cancer have made great progress, but the diagnosis and treatment of breast cancer have made great progress. How to effectively control the progress of breast cancer, prolong the survival period of the patients with breast cancer, prevent or reduce the recurrence of breast cancer and reduce the mortality of breast cancer is still a difficult problem for the treatment of breast cancer. Currently, biological targeting therapy has been one of the conventional treatment methods after traditional surgical treatment, radiotherapy and chemotherapy. McAbs and small molecular targeting drugs have been widely used in various cancers including breast cancer, and it has been proved to be effective for many kinds of cancers including breast cancer. However, single target therapy still exists that the tumor is prone to mutation and produces drug resistance, resulting in the occurrence of tumor treatment failure. The combination of two or more targeted drugs has a potential application prospect to improve the therapeutic effect of many kinds of tumors, including breast cancer. Therefore, it is one of the key factors for the combination of targeted drugs to treat a variety of tumors such as breast cancer.
Previous studies have shown that EGFR plays an important role in the development of a variety of tumors, especially the overexpression of.EGFR, a malignant tumor, such as head and neck cancer, lung cancer, and other epithelial neoplasm, is important for the occurrence, development, prognosis and so on of the malignant tumor. For EGFR targeted drugs, such as AI Bi, and Tai Xin Sheng The clinical application of small molecular targeting drug yyesa and other clinical applications all effectively inhibit the growth and metastasis of epithelial derived malignant tumor tissues. In the course of the development of breast cancer, EGFR also plays an important role. It has been proved that the activation of EGFR on the surface of breast cancer cells leads to the activation of intracellular tyrosine kinase area. In addition, overexpression of EGFR in tissue can cause the continuous activation of multiple signal transduction pathways, promote protein synthesis, inhibit cell apoptosis, accelerate cell division and proliferation, and promote the occurrence and development of breast cancer. Therefore, EGFR plays an important role in the occurrence, development and prognosis of breast cancer. It is the treatment of breast cancer. One of the targets.
The role of indolamine 2,3- dioxygenase (indoleamine2,3-dioxygenase, IDO) in human and animal metabolism has attracted more and more attention, and widely distributed in human and other mammalian tissues other than the liver. IDO through the degradation of tryptophan, the local tryptophan deficiency and the proliferation of T cells are suppressed. In addition, I DO degradation product, canine urea and its derivatives, also participate in the immunoregulation process to induce apoptosis of T cells. The study shows that the high expression of IDO is closely related to the tumor immune escape in many tumors, including breast cancer. Therefore, IDO is one of the targets for a variety of tumours, including breast cancer. It shows that IDO inhibitors play a role by inhibiting IDO to restore the proliferation of T cells to normal state, and thus have a certain therapeutic effect on inhibiting tumor invasion and metastasis. At present, the tumor biological targeting therapy is gradually going deep, from the treatment of single target to multiple targets and in a variety of methods. In fact, multi target combination therapy has many advantages, such as reducing drug use and reducing the occurrence of tumor resistance. Therefore, multi target combination therapy has become one of the trends in the future development of cancer treatment. Although EGFR and IDO are reported in breast cancer, the combined application of EGFR and IDO two inhibitors has not been seen. The development of prebreast cancer and the progress of prognosis are studied. This article is to detect the co expression of EGFR and IDO in breast cancer, and to jointly inhibit EGFR and IDO, and provide a new therapeutic strategy for exploring the combination of multiple targets and the prognosis of breast cancer.
The distribution of co expression of EGFR and IDO in breast cancer patients was first detected to lay the foundation for the combined inhibition of EGFR and IDO double targets and the combined inhibition of the development prognosis of breast cancer. 110 cases of breast cancer specimens were made by paraffin section and detected by immunohistochemical method. The results confirmed that there was indeed a total of EGFR and IDO in breast cancer tissues. The total expression rate of the two cases was a certain proportion in the patients with breast cancer. The total expression rate was 29.1%, the synchronic co expression of 19.1%, the difference co expressed 10%., but the ultrastructural expression regions of the two were different. For the IDO, the cytoplasm of the malignant ductal cells was mostly not detected in the adjacent basal cells. The degree of synchronous co expression of.IDO and EGFR on the membrane of EGFR mainly expressed in epithelial cells and basal cells was low to high. At the same time, there was also a difference co expression between IDO and EGFR, such as IDO low expression, moderate or high expression of EGFR, while EGFR low expression, IDO moderate or high expression. We tested 8 kinds of breast cancer cell lines, using immunocytochemistry, RT-PCR and Westen Blot methods to verify each other. The results showed that the co expression of IDO and EGFR was also present in the breast cancer cell lines. The above two experimental results confirmed that the prerequisite for the combined treatment of breast cancer with EGFR and IDO was established. On the other hand, by using in vitro cell test, we used Abbey, Iressa and 1- methyl tryptophan. It was proved that the combined inhibition of the expression of EGFR and IDO could effectively inhibit the growth and invasion and metastasis of breast cancer cells, which laid the foundation for our future research in vivo.
Conclusion:
1, our study confirms for the first time that there is a co expression of EGFR and IDO in breast cancer patients, and there are two cases of differential co expression and synchronous co expression.
2, our study confirms for the first time that co expression of EGFR and IDO is also present in breast cancer cell lines.
3, our in vitro experiments confirmed that the combined inhibition of EGFR and IDO could effectively inhibit the growth, infiltration and metastasis of breast cancer cells, and obviously better than the individual inhibition. It showed that the synergistic effects of the two were combined.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.9

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