褪黑素协同5-FU在结肠癌中作用与分子机制研究

发布时间：2018-07-02 13:15

本文选题：褪黑素 + 5-FU　；参考：《大连医科大学》2017年硕士论文

【摘要】：目的:结肠癌在全球范围内其死亡率高居第三位。5-FU在结肠癌治疗中是最常用的化疗药物之一,但是由于其本身的致毒性导致其治疗指数的局限性。褪黑素已被证实具有抗肿瘤活性并且可与其他化学治疗药联合用于癌症治疗。然而,尚未报道褪黑素与5-FU联合用药治疗结肠癌能否提高5-FU的治疗疗效。我们的研究目的是为了探究这样一种联合用药褪黑素是否可协同提高5-FU抑制结肠癌细胞增殖和促进凋亡作用。方法:本研究中,单独褪黑素、单独5-FU和褪黑素联合5-FU处理人结肠癌细胞SW620和LOVO,MTT法检测细胞活力,通过克隆形成实验、划痕实验、侵袭实验、凋亡和周期实验检测不同药物处理的细胞生物学功能。在分子机制研究中,Western blot检测PI3K/AKT/NF-κB/iNOS信号通路的一些关键蛋白。进一步通过不同药物处理组进行激光共聚焦扫描显微镜和染色质免疫共沉淀实验研究NF-κB p50/p65的核定位及它们与iNOS的启动子区的结合情况。最后,利用荷瘤小鼠实验,研究褪黑素协同增强5-FU的抗肿瘤活性。结果:与单独5-FU药物处理组相比,联合褪黑素与5-FU药物处理组能够显著抑制结肠癌细胞增殖、克隆形成、细胞迁移及侵袭并能够显著改变细胞的形态。除此之外,联合褪黑素和5-FU药物处理组能够更明显的诱导结肠癌细胞周期阻滞和激活细胞凋亡蛋白酶依赖的凋亡通路。进一步的分子机制实验表明这样一种联合用药与单独5-FU药物处理更能显著抑制PI3K/AKT和NF-κB/iNOS信号通路的激活。褪黑素协同加强5-FU抑制PI3K或iNOS的活性。更重要的是,与单独5-FU药物处理组相比,联合褪黑素的5-FU药物处理组更加显著地抑制了荷瘤小鼠模型体内的结肠癌的生长,同样地更加明显抑制了AKT和iNOS的活性。结论:本研究发现与单独5-FU药物处理组相比,联合褪黑素的5-FU药物处理组能够显著提高抑制结肠癌生长作用。更重要的是,研究表明这样一种在结肠癌中抗增殖和促凋亡的作用提高是通过调节多个信号通路包括PI3K/AKT和NF-κB/iNOS。
[Abstract]:Objective: colon cancer is one of the most commonly used chemotherapeutic drugs in colon cancer treatment, but its treatment index is limited due to its toxicity. Melatonin has been proven to have anti-tumor activity and can be used in combination with other chemotherapeutic drugs for cancer treatment. However, whether the combination of melatonin and 5-FU can improve the efficacy of 5-FU in colon cancer has not been reported. Our aim was to investigate whether melatonin, a combination of these drugs, can increase the proliferation and apoptosis of colon cancer cells in combination with 5-FU. Methods: in this study, single melatonin, 5-FU and 5-FU combined with 5-FU were used to detect the viability of human colon cancer cell line SW620 and LOVOA MTT. Apoptosis and periodic experiments were used to detect the biological function of different drugs. Some key proteins in the PI3K / AKT / NF- 魏 B / iNOS signaling pathway were detected by Western blot. Furthermore, laser confocal scanning microscopy and chromatin immunoprecipitation were used to study the nuclear localization of NF- 魏 B p50/p65 and the binding of NF- 魏 B p50/p65 to the promoter region of iNOS. Finally, the anti-tumor activity of 5-Fu was studied by using melatonin in tumor-bearing mice. Results: the combination of melatonin and 5-FU could significantly inhibit the proliferation, clone formation, cell migration and invasion of colon cancer cells and significantly change the morphology of colon cancer cells. In addition, the combination of melatonin and 5-FU could induce cell cycle arrest and activation of apoptotic protease-dependent apoptosis pathway in colon cancer cells. Further molecular mechanism experiments showed that the combination of 5-FU and 5-FU could significantly inhibit the activation of PI3K / AKT and NF- 魏 B / iNOS signaling pathway. Melatonin synergistically inhibits the activity of PI3K or iNOS by 5-FU. More importantly, 5-FU combined with melatonin significantly inhibited the growth of colon cancer and the activity of AKT and iNOS in tumor-bearing mice compared with 5-FU alone. Conclusion: compared with 5-FU alone, 5-FU combined with melatonin can significantly inhibit the growth of colon cancer. More importantly, studies have shown that such an anti-proliferation and pro-apoptotic role in colon cancer is enhanced by regulating multiple signaling pathways, including PI3K / AKT and NF- 魏 B / iNOS.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35

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