联合近红外荧光分子肿瘤靶向探针的热效应与免疫因子协同治疗甲状腺肿瘤的研究
发布时间：2018-11-23 19:34
【摘要】：甲状腺癌是内分泌系统最常见的恶性肿瘤,且其发病率在全世界均呈逐年增加趋势。临床上,甲状腺癌对外照射放疗和化疗均不敏感,传统的治疗以器官损毁性手术为主,病人面临终身药物替代和严重手术并发症的风险。近年来,热疗成为恶性肿瘤综合治疗的新兴力量,在甲状腺癌的治疗方面也有学者进行了探索,尤其针对甲状腺微小癌和侧颈部淋巴结复发的热消融治疗。热消融与传统手术或放化疗相比,优势在于微创、全身毒性低,另外热疗时肿瘤细胞产生的热休克蛋白70(HSP70)可呈递肿瘤抗原,激活Toll样受体4(Toll-like receptors-4,TLR-4)从而增强机体的抗肿瘤免疫。但是,热消融治疗也存在一些亟待解决的问题,一方面临床上由影像学介导的热能治疗较为粗放,实现肿瘤治疗的彻底性具有一定缺陷,另一方面,HSP70作为分子伴侣能保护细胞,减少高热引起的凋亡,从而降低高热对肿瘤杀伤效果。本研究针对上述问题,尝试联合应用靶向高热疗法来治疗甲状腺癌。论文构建了AG修饰的近红外染料探针AG-IR820,将其靶向于甲状腺癌细胞,通过近红外线照射产热杀伤癌细胞。同时实验中联合使用槲皮素(Quercetin)来抑制HSP70的表达、应用脂多糖(lipopolysaccharide,LPS)激活TLR-4通路增强抗肿瘤免疫。结果显示:利用AG修饰IR820能明显提高其细胞摄取能力和肿瘤的靶向能力;在体外和体内实验中,三联靶向热疗与单独疗法相比有更强的抑制肿瘤效果。本研究表明这种新的联合治疗策略能显著改善热疗效果,有可能成为甲状腺癌治疗的新方法。本论文主要分为六个部分,具体如下:1.绪论介绍本文的研究背景及设计思路。2.肿瘤靶向探针AG-IR820的合成与表征结论:AG-IR820制备成功,分子量1105.34,与理论计算结果一致;AG-IR820吸收光谱峰值691nm,发射光谱峰值823nm,与IR820荧光特性一致。3.AG-IR820的肿瘤靶向能力研究结论:AG的修饰作用使IR820具备了肿瘤靶向能力,AG-IR820能被人甲状腺癌TT细胞系特异性摄取,在荷瘤裸鼠体内可靶向定位到肿瘤病灶,且有较快速的浓聚速度(小于4h)和较长的滞留时间(超过24h)。4.AG-IR820的热疗性能研究结论:AG-IR820和IR820在5μM浓度时对TT细胞无明显毒性。在近红外光照射时,AG-IR820产热使温度上升至43℃,具有杀伤肿瘤细胞的能力,且比IR820热疗效果更明显。5.AG-IR820+Quercetin+LPS联合应用的体外实验研究结论:体外实验中,Quercetin在100μM浓度时可有效减少HSP70的表达而没有明显毒性;2μg/m L的脱毒LPS无明显的细胞毒性。AG-IR820介导的热疗联合Quercetin能提高热疗的杀伤能力,再联合脱毒LPS,可激活巨噬细胞释放肿瘤TNF-α,进一步增强热疗效果。6.AG-IR820+Quercetin+LPS联合应用的体内实验研究结论:AG-IR820+Quercetin+LPS的三联疗法对人甲状腺TT细胞系裸鼠移植肿瘤模型的治疗效果要明显优于其他单独或二联疗法。联合HSP70抑制剂和TLR-4激活剂可以增强热疗效果。
[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，
本文编号：2352494
[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，
本文编号：2352494
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