本文選題：EGFR + IDO��； 參考：《吉林大學(xué)》2015年博士論文

【摘要】：乳腺癌是女性最常見的惡性腫瘤，近年來，全世界乳腺癌發(fā)病率不斷增高，在我國乳腺癌也呈現(xiàn)不斷增高的趨勢。尤其值得關(guān)注的是乳腺癌目前已成為導(dǎo)致女性死亡的重要因素之一。盡管乳腺癌的診斷與治療已經(jīng)取得了很大進(jìn)展，但是如何有效地控制乳腺癌的進(jìn)展，延長乳腺癌患者的生存期以及防止或減少乳腺癌的復(fù)發(fā)，降低乳腺癌的死亡率仍是目前乳腺癌治療的難題。目前，生物靶向治療已經(jīng)是繼傳統(tǒng)的手術(shù)治療、放療及化療后的常規(guī)治療手段之一。包括人源化單抗及小分子靶向藥物已經(jīng)在包括乳腺癌在內(nèi)的多種癌癥治療當(dāng)中廣泛應(yīng)用，并且證實(shí)對包括乳腺癌在內(nèi)的多種癌癥均有一定的療效。但是單一靶點(diǎn)治療仍存在腫瘤易發(fā)生變異并產(chǎn)生耐藥性，從而導(dǎo)致腫瘤治療失效的現(xiàn)象發(fā)生。如何采用兩種或多種靶向藥物聯(lián)合治療對于提高包括乳腺癌等多種腫瘤的治療效果具有潛在的應(yīng)用前景。因此，尋找可聯(lián)合應(yīng)用于腫瘤生物治療的靶點(diǎn)是聯(lián)合應(yīng)用靶向藥物治療乳腺癌等多種腫瘤的關(guān)鍵因素之一。 以往的研究表明，EGFR在多種腫瘤的發(fā)生、發(fā)展中具有重要作用，尤其是對于頭頸癌、肺癌等多種上皮組織源性的惡性腫瘤。EGFR的過表達(dá)對于上述惡性腫瘤的發(fā)生、發(fā)展、預(yù)后轉(zhuǎn)歸等均具有重要作用，針對EGFR靶向治療藥物如愛必妥、泰欣生以及小分子靶向藥物易瑞沙等的臨床應(yīng)用均有效地抑制了上皮源性的惡性腫瘤組織的生長與轉(zhuǎn)移。在乳腺癌的發(fā)生發(fā)展過程中，EGFR也同樣發(fā)揮了重要作用。研究證實(shí)，存在于乳腺癌細(xì)胞表面EGFR的活化，導(dǎo)致胞內(nèi)酪氨酸激酶區(qū)的活化。另外，EGFR在組織中的過度表達(dá)可造成多條信號傳導(dǎo)通路的持續(xù)性激活，促進(jìn)蛋白質(zhì)合成，抑制細(xì)胞的凋亡，加速細(xì)胞的分裂和增殖，從而促進(jìn)乳腺癌的發(fā)生、發(fā)展。因此，EGFR對于乳腺癌的發(fā)生、發(fā)展及預(yù)后轉(zhuǎn)歸均起著重要作用，是乳腺癌治療的靶點(diǎn)之一。 吲哚胺2，3-雙加氧酶（indoleamine2，3-dioxygenase，IDO）在人及動(dòng)物代謝中的作用越來越引起研究者的廣泛重視，并且廣泛分布于人和其他哺乳動(dòng)物除肝臟以外的組織中， IDO通過降解色氨酸，使局部造成色氨酸缺乏進(jìn)而T細(xì)胞增殖受到抑制。另外，，IDO的降解產(chǎn)物犬尿素及其衍生物也參與免疫調(diào)節(jié)過程可誘導(dǎo)T細(xì)胞凋亡。研究顯示，IDO的高表達(dá)在包括乳腺癌在內(nèi)的多種腫瘤中與腫瘤發(fā)生免疫逃逸密切相關(guān)，因此，IDO是包括乳腺癌在內(nèi)的多種腫瘤的有研究前景的靶點(diǎn)之一。許多研究報(bào)道顯示，利用IDO的抑制劑通過抑制IDO發(fā)揮作用，使T細(xì)胞的增殖恢復(fù)到正常狀態(tài)，從而對抑制腫瘤的浸潤轉(zhuǎn)移具有一定的治療作用。目前，腫瘤生物靶向治療也逐漸走向深入，從單一靶點(diǎn)的治療到多靶點(diǎn)以及多種手段聯(lián)合治療,并且實(shí)踐證實(shí),多靶點(diǎn)聯(lián)合治療具有減少藥物用量，減少腫瘤耐藥性發(fā)生等諸多優(yōu)勢。因此，多靶點(diǎn)聯(lián)合腫瘤治療已經(jīng)成為腫瘤治療未來發(fā)展的趨勢之一。盡管EGFR和IDO在乳腺癌中的研究都分別有報(bào)道，但目前尚未見到EGFR和IDO二者抑制劑聯(lián)合應(yīng)用干預(yù)乳腺癌的發(fā)生發(fā)展、預(yù)后轉(zhuǎn)歸進(jìn)程的相關(guān)研究。本文擬通過檢測乳腺癌中EGFR與IDO共表達(dá)的情況，從而聯(lián)合抑制EGFR與IDO，為探討多靶點(diǎn)聯(lián)合抑制乳腺癌的發(fā)展及預(yù)后提供新的治療策略。 首先檢測乳腺癌患者中EGFR與IDO共表達(dá)的分布情況，為實(shí)施聯(lián)合抑制EGFR與IDO雙靶點(diǎn)，從而聯(lián)合抑制乳腺癌的發(fā)展預(yù)后奠定基礎(chǔ)。通過對110例乳腺癌患者標(biāo)本制成石蠟切片，用免疫組化方法檢測，結(jié)果證實(shí)乳腺癌組織中確實(shí)存在EGFR和IDO共表達(dá)情況，并且二者共表達(dá)情況在乳腺癌患者中占有一定的比例。總共表達(dá)率為29.1%，同步共表達(dá)19.1%，差異共表達(dá)10%。但二者的超微結(jié)構(gòu)表達(dá)區(qū)域是有差異的。對于IDO，主要表達(dá)在惡性導(dǎo)管細(xì)胞的細(xì)胞漿，在臨近的基底細(xì)胞大多檢測不到。對于EGFR主要表達(dá)在上皮細(xì)胞和基底細(xì)胞的細(xì)胞膜上。IDO和EGFR的同步共表達(dá)程度由低到高。同時(shí)，IDO和EGFR也存在差異共表達(dá)，如IDO低表達(dá)時(shí)，EGFR中度或高表達(dá)。而EGFR低表達(dá)時(shí)，IDO中度或高表達(dá)。為進(jìn)一步通過試驗(yàn)研究證實(shí)上述實(shí)驗(yàn)結(jié)果，我們對8種乳腺癌細(xì)胞系進(jìn)行了檢測，利用免疫細(xì)胞化學(xué)，RT-PCR及Westen Blot的方法相互驗(yàn)證。結(jié)果表明，乳腺癌細(xì)胞系中也同樣存在IDO和EGFR共表達(dá)情況。上述兩方面試驗(yàn)結(jié)果證實(shí)開展以EGFR和IDO聯(lián)合治療乳腺癌的前提條件是成立的。在此基礎(chǔ)上，我們通過體外細(xì)胞試驗(yàn)采用艾比妥、易瑞沙、1-甲基色氨酸聯(lián)合用藥，初步證實(shí)聯(lián)合抑制EGFR和IDO的表達(dá)能夠有效地抑制乳腺癌細(xì)胞的生長與侵襲浸潤及轉(zhuǎn)移，這為我們今后開展體內(nèi)試驗(yàn)研究奠定了基礎(chǔ)。 結(jié)論： 1、我們的研究首次證實(shí)乳腺癌患者中存在EGFR和IDO的共表達(dá)情況，并且存在差異共表達(dá)和同步共表達(dá)兩種情況。 2、我們的研究首次證實(shí)乳腺癌細(xì)胞系中也同樣存在EGFR和IDO的共表達(dá)情況。 3、我們的體外試驗(yàn)證實(shí)通過對EGFR和IDO聯(lián)合抑制，能有效地抑制乳腺癌細(xì)胞的生長、浸潤與轉(zhuǎn)移，并且明顯優(yōu)于單獨(dú)抑制情況。表明二者聯(lián)合抑制存在協(xié)同效應(yīng)。
[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.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R737.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Ozlem Yersa L;Sabri Barutca;;Biological subtypes of breast cancer: Prognostic and therapeutic implications[J];World Journal of Clinical Oncology;2014年03期

本文編號：1856250