纳米金改善结肠癌细胞的耐药性及其机制的研究

发布时间：2018-05-10 06:11

  本文选题：纳米金 + 奥沙利铂　； 参考：《暨南大学》2016年硕士论文

【摘要】：目的：观察纳米金(gold nanoparticles, GNPs)对奥沙利铂(Oxaliplatin, L-OHP)抗耐药结肠癌细胞(COLO 205/L-OHP)的影响并初步探讨其相关机制。方法：首先根据化学合成法制备纳米金,并用紫外-可见吸收光谱和透射电子显微镜鉴定。实验第一步通过MTT比色法、PCR、Western blot分别检测耐药结肠癌细胞COLO 205/L-OHP对药物L-OHP的敏感程度、MDR和MRP1基因表达以及P-gp蛋白的表达,并与正常结肠癌细胞COLO 205/S对比；第二步将纳米金联合奥沙利铂对COLO 205/L-OHP进行处理,采用MTT法和流式细胞术分别检测增殖抑制率和凋亡率；第三步用不同浓度的纳米金处理COLO 205/L-OHP,通过ELISA检测细胞培养液内b-FGF的浓度,Western blot检测细胞内b-FGF、Bcl-2蛋白的表达量。结果：纳米金的最大吸收峰在520 nm处,粒径约10 nm大小；在浓度为1.5μg/mL的L-OHP中,COLO 205/L-OHP细胞增殖抑制率明显低于COLO 205/S(P0.05)；与COLO 205/S目比,COLO 205/L-OHP耐药基因MDR、MRP1以及P-gp表达增多；联合用药组COLO 205/L-OHP细胞增殖抑制率、凋亡率高于单用奥沙利铂组(P0.05)。COLO 205/L-OHP培养液内b-FGF以及细胞内所表达的b-FGF、Bcl-2含量与COLO 205/S相比明显增高(P0.05),单纯纳米金对COLO 205/L-OHP细胞增殖无抑制作用(P0.05),但是经纳米金处理后,COLO 205/L-OHP细胞中b-FGF和Bcl-2含量明显减少,且具有浓度依赖性。结论：纳米金能够降低COLO 205/L-OHP细胞内b-FGF含量,使细胞内抗凋亡蛋白表达减少,增加奥沙利铂毒性作用,改善耐药情况。
[Abstract]:Aim: to investigate the effect of nanoparticles (GNPs) on oxaliplatin (L-OHP-) -resistant colon cancer cell line (COLO _ 205 / L-OHP) and its related mechanism. Methods: gold nanoparticles were prepared by chemical synthesis method and identified by UV-vis absorption spectrum and transmission electron microscope. In the first step of the experiment, the sensitivity of COLO 205/L-OHP to drug L-OHP and the expression of P-gp protein were detected by MTT colorimetric assay and Western blot, respectively, and compared with those of normal colon cancer cell line COLO 205 / S. In the second step, COLO 205/L-OHP was treated with gold nanoparticles combined with oxaliplatin. Proliferation inhibition rate and apoptosis rate were detected by MTT and flow cytometry. In the third step, COLO 205 / L -OHP was treated with different concentrations of nano-gold, and the expression of b-FGF- Bcl 2 protein was detected by ELISA, and the concentration of b-FGF in cell culture medium and the expression of b-FGF- Bcl 2 protein were detected by Western blot. Results: the maximum absorption peak of gold nanoparticles was at 520nm and the diameter was about 10nm. The inhibition rate of COLO 205/L-OHP cell proliferation in 1.5 渭 g/mL L-OHP was significantly lower than that of COLO 205% SNP 0.05, and the expression of COLO 205/L-OHP resistance gene MRP1 and P-gp was higher than that of COLO 205S. The proliferation inhibition rate of COLO 205/L-OHP cells in combination group, The rate of apoptosis was higher than that of oxaliplatin alone in b-FGF and b-FGFG Bcl 2 expression in the culture medium of P0.05N 路Colo 205/L-OHP and COLO 205 / S. The effect of Nano Gold alone on the proliferation of COLO 205/L-OHP cells was not inhibited (P0.05N), but the expression of b-FGFGF-Bcl 2 was significantly higher than that of COLO 205S. However, after the treatment of nanocrystalline gold, COLO 205/L-OHP was not inhibited. The contents of b-FGF and Bcl-2 in the cells decreased significantly. And it is concentration-dependent. Conclusion: gold nanoparticles can decrease the content of b-FGF in COLO 205/L-OHP cells, decrease the expression of anti-apoptotic protein, increase oxaliplatin toxicity and improve drug resistance.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.35
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本文编号：1868137