【摘要】：甲狀腺癌是內(nèi)分泌系統(tǒng)最常見的惡性腫瘤,且其發(fā)病率在全世界均呈逐年增加趨勢(shì)。臨床上,甲狀腺癌對(duì)外照射放療和化療均不敏感,傳統(tǒng)的治療以器官損毀性手術(shù)為主,病人面臨終身藥物替代和嚴(yán)重手術(shù)并發(fā)癥的風(fēng)險(xiǎn)。近年來,熱療成為惡性腫瘤綜合治療的新興力量,在甲狀腺癌的治療方面也有學(xué)者進(jìn)行了探索,尤其針對(duì)甲狀腺微小癌和側(cè)頸部淋巴結(jié)復(fù)發(fā)的熱消融治療。熱消融與傳統(tǒng)手術(shù)或放化療相比,優(yōu)勢(shì)在于微創(chuàng)、全身毒性低,另外熱療時(shí)腫瘤細(xì)胞產(chǎn)生的熱休克蛋白70(HSP70)可呈遞腫瘤抗原,激活Toll樣受體4(Toll-like receptors-4,TLR-4)從而增強(qiáng)機(jī)體的抗腫瘤免疫。但是,熱消融治療也存在一些亟待解決的問題,一方面臨床上由影像學(xué)介導(dǎo)的熱能治療較為粗放,實(shí)現(xiàn)腫瘤治療的徹底性具有一定缺陷,另一方面,HSP70作為分子伴侶能保護(hù)細(xì)胞,減少高熱引起的凋亡,從而降低高熱對(duì)腫瘤殺傷效果。本研究針對(duì)上述問題,嘗試聯(lián)合應(yīng)用靶向高熱療法來治療甲狀腺癌。論文構(gòu)建了AG修飾的近紅外染料探針AG-IR820,將其靶向于甲狀腺癌細(xì)胞,通過近紅外線照射產(chǎn)熱殺傷癌細(xì)胞。同時(shí)實(shí)驗(yàn)中聯(lián)合使用槲皮素(Quercetin)來抑制HSP70的表達(dá)、應(yīng)用脂多糖(lipopolysaccharide,LPS)激活TLR-4通路增強(qiáng)抗腫瘤免疫。結(jié)果顯示:利用AG修飾IR820能明顯提高其細(xì)胞攝取能力和腫瘤的靶向能力;在體外和體內(nèi)實(shí)驗(yàn)中,三聯(lián)靶向熱療與單獨(dú)療法相比有更強(qiáng)的抑制腫瘤效果。本研究表明這種新的聯(lián)合治療策略能顯著改善熱療效果,有可能成為甲狀腺癌治療的新方法。本論文主要分為六個(gè)部分,具體如下:1.緒論介紹本文的研究背景及設(shè)計(jì)思路。2.腫瘤靶向探針AG-IR820的合成與表征結(jié)論:AG-IR820制備成功,分子量1105.34,與理論計(jì)算結(jié)果一致;AG-IR820吸收光譜峰值691nm,發(fā)射光譜峰值823nm,與IR820熒光特性一致。3.AG-IR820的腫瘤靶向能力研究結(jié)論:AG的修飾作用使IR820具備了腫瘤靶向能力,AG-IR820能被人甲狀腺癌TT細(xì)胞系特異性攝取,在荷瘤裸鼠體內(nèi)可靶向定位到腫瘤病灶,且有較快速的濃聚速度(小于4h)和較長(zhǎng)的滯留時(shí)間(超過24h)。4.AG-IR820的熱療性能研究結(jié)論:AG-IR820和IR820在5μM濃度時(shí)對(duì)TT細(xì)胞無明顯毒性。在近紅外光照射時(shí),AG-IR820產(chǎn)熱使溫度上升至43℃,具有殺傷腫瘤細(xì)胞的能力,且比IR820熱療效果更明顯。5.AG-IR820+Quercetin+LPS聯(lián)合應(yīng)用的體外實(shí)驗(yàn)研究結(jié)論:體外實(shí)驗(yàn)中,Quercetin在100μM濃度時(shí)可有效減少HSP70的表達(dá)而沒有明顯毒性;2μg/m L的脫毒LPS無明顯的細(xì)胞毒性。AG-IR820介導(dǎo)的熱療聯(lián)合Quercetin能提高熱療的殺傷能力,再聯(lián)合脫毒LPS,可激活巨噬細(xì)胞釋放腫瘤TNF-α,進(jìn)一步增強(qiáng)熱療效果。6.AG-IR820+Quercetin+LPS聯(lián)合應(yīng)用的體內(nèi)實(shí)驗(yàn)研究結(jié)論:AG-IR820+Quercetin+LPS的三聯(lián)療法對(duì)人甲狀腺TT細(xì)胞系裸鼠移植腫瘤模型的治療效果要明顯優(yōu)于其他單獨(dú)或二聯(lián)療法。聯(lián)合HSP70抑制劑和TLR-4激活劑可以增強(qiáng)熱療效果。
[Abstract]:Thyroid carcinoma is the most common malignant tumor in the endocrine system, and its incidence is increasing year by year in the world. Clinically, thyroid cancer is insensitive to external irradiation, radiotherapy and chemotherapy. The traditional treatment is mainly organ lesion surgery. Patients face the risk of life-long drug substitution and severe complications. In recent years, hyperthermia has become a new force in the comprehensive treatment of malignant tumors. Some scholars have also explored the treatment of thyroid cancer, especially for the treatment of thyroid microcarcinoma and the recurrence of lateral cervical lymph nodes. Compared with conventional surgery or radiotherapy and chemotherapy, thermal ablation has the advantages of minimally invasive and low systemic toxicity. In addition, heat shock protein 70 (HSP70) produced by tumor cells during hyperthermia can present tumor antigens and activate Toll like receptor 4 (Toll-like receptors-4,). TLR-4) thus enhances the body's anti-tumor immunity. However, there are some urgent problems in thermal ablation therapy. On the one hand, thermal energy therapy mediated by imaging is relatively extensive, which has certain defects to achieve complete tumor treatment, on the other hand, HSP70 as a molecular chaperone can protect cells, reduce apoptosis induced by high fever, and thus reduce the killing effect of high fever on tumor. In order to solve the above problems, this study attempted to use targeted hyperthermia therapy to treat thyroid cancer. In this paper, AG modified near infrared dye probe AG-IR820, was constructed to target thyroid cancer cells and kill cancer cells by near infrared irradiation. Quercetin (Quercetin) was used to inhibit the expression of HSP70 and lipopolysaccharide (lipopolysaccharide,LPS) was used to activate TLR-4 pathway to enhance anti-tumor immunity. The results showed that AG modified IR820 could significantly improve the ability of cell uptake and tumor targeting, and in vitro and in vivo, triple targeted hyperthermia had a stronger effect on tumor inhibition than that of single therapy. This study suggests that this new combined therapy strategy can significantly improve the effect of hyperthermia and may become a new method for thyroid cancer treatment. This paper is divided into six parts, as follows: 1. Introduction introduces the research background and design ideas of this paper. 2. Synthesis and characterization of tumor targeting probe AG-IR820 conclusion: the molecular weight of AG-IR820 was 1105.34, which was consistent with the theoretical results. The peak value of AG-IR820 absorption spectrum was 691 nm, and the peak value of emission spectrum was 823 nm, which was consistent with the fluorescence characteristics of IR820. Conclusion: the modification of AG makes IR820 possess tumor targeting ability. AG-IR820 can be specifically ingested by human thyroid carcinoma TT cell line, and can be targeted to the tumor focus in nude mice. The hyperthermia properties of 4.AG-IR820 were studied. Conclusion: AG-IR820 and IR820 have no obvious toxicity to TT cells at 5 渭 M concentration. When irradiated by near-infrared light, the heat production of AG-IR820 rises to 43 鈩，

本文編號(hào)：2352494