本文選題：納米金 + 奧沙利鉑��； 參考：《暨南大學(xué)》2016年碩士論文

【摘要】：目的：觀察納米金(gold nanoparticles, GNPs)對奧沙利鉑(Oxaliplatin, L-OHP)抗耐藥結(jié)腸癌細(xì)胞(COLO 205/L-OHP)的影響并初步探討其相關(guān)機(jī)制。方法：首先根據(jù)化學(xué)合成法制備納米金,并用紫外-可見吸收光譜和透射電子顯微鏡鑒定。實(shí)驗(yàn)第一步通過MTT比色法、PCR、Western blot分別檢測耐藥結(jié)腸癌細(xì)胞COLO 205/L-OHP對藥物L(fēng)-OHP的敏感程度、MDR和MRP1基因表達(dá)以及P-gp蛋白的表達(dá),并與正常結(jié)腸癌細(xì)胞COLO 205/S對比；第二步將納米金聯(lián)合奧沙利鉑對COLO 205/L-OHP進(jìn)行處理,采用MTT法和流式細(xì)胞術(shù)分別檢測增殖抑制率和凋亡率；第三步用不同濃度的納米金處理COLO 205/L-OHP,通過ELISA檢測細(xì)胞培養(yǎng)液內(nèi)b-FGF的濃度,Western blot檢測細(xì)胞內(nèi)b-FGF、Bcl-2蛋白的表達(dá)量。結(jié)果：納米金的最大吸收峰在520 nm處,粒徑約10 nm大小；在濃度為1.5μg/mL的L-OHP中,COLO 205/L-OHP細(xì)胞增殖抑制率明顯低于COLO 205/S(P0.05)；與COLO 205/S目比,COLO 205/L-OHP耐藥基因MDR、MRP1以及P-gp表達(dá)增多；聯(lián)合用藥組COLO 205/L-OHP細(xì)胞增殖抑制率、凋亡率高于單用奧沙利鉑組(P0.05)。COLO 205/L-OHP培養(yǎng)液內(nèi)b-FGF以及細(xì)胞內(nèi)所表達(dá)的b-FGF、Bcl-2含量與COLO 205/S相比明顯增高(P0.05),單純納米金對COLO 205/L-OHP細(xì)胞增殖無抑制作用(P0.05),但是經(jīng)納米金處理后,COLO 205/L-OHP細(xì)胞中b-FGF和Bcl-2含量明顯減少,且具有濃度依賴性。結(jié)論：納米金能夠降低COLO 205/L-OHP細(xì)胞內(nèi)b-FGF含量,使細(xì)胞內(nèi)抗凋亡蛋白表達(dá)減少,增加奧沙利鉑毒性作用,改善耐藥情況。
[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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R735.35
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本文編號：1868137