氟尿嘧啶联合姜黄素纳米剂型抑制肝癌细胞的研究  

Co-delivery of fluorouracil and curcumin by nano-layered double hydroxide for inhibition of hepatocellular carcinoma cells in vitro

作　　者：董伟[1] 刘福晨 倪俊声[1] 李鹏鹏[1] 郭兴刚 刘辉[1] 

机构地区：[1]第二军医大学东方肝胆外科医院肝外三科,上海200438

出　　处：《中国现代医学杂志》2016年第9期16-22,共7页China Journal of Modern Medicine

基　　金：国家自然科学基金面上项目(No:NSFC81271694);科技部国际合作专项(No:010S2012ZR0058;2011DFA32980)

摘　　要：目的探究纳米层状双氢氧化物(LDH)共载氟尿嘧啶(Fu)姜黄素(Cur)混合剂型对肝癌细胞的抑制作用。方法通过共沉淀法合成纳米LDH混合剂型,并对其进行详细表征,如电镜(TEM)、X射线衍射(XRD)及动态光散射技术(DLS)检测,细胞计数试剂盒(CCK8)测定Fu组、Fu+Cur组及纳米剂型组对7721、Hep G2和LM3细胞的增殖抑制作用,同时凋亡试剂盒检测各组对7721细胞的凋亡效应,Western blot检测各组对细胞抗凋亡相关基因Bcl-2蛋白的表达及下游Caspase的活化情况。结果合成的纳米LDH混合剂型具有较好的均一性,DLS检测提示其粒径约为400nm,XRD提示Fu及Cur通过水平方式插入LDH层间;CCK8结果提示,相对于单纯的Fu组、Fu+Cur组和LDH-Fu组,LDH-Fu-Cur组对LM3、Hep G2和7721细胞具有更强的增殖抑制作用(P<0.05)。相对于单纯的Fu组、Fu+Cur组和LDH-Fu组,LDH-Fu-Cur组对LM3、Hep G2和7721细胞具有更强的促凋亡效应作用(P<0.05),35μg/ml浓度的Fu纳米混合剂型组凋亡率高达(87.0±4.7)%,而单纯Fu仅为(23.0±2.3)%;Western blot检测结果显示,纳米混合剂型能够下调7721细胞Bcl-2水平。结论纳米LDH混合剂型具有良好的抗肿瘤效应,其机制可能为下调肿瘤抗凋亡基因Bcl-2从而引起下游Caspase活化。Objective To evaluate the effects of fluorouracil and curcumin co-loaded nano-layered double hydroxide（NLDH） on proliferation and apoptosis of hepatocellular carcinoma cell lines. Methods NLDH was synthesized by co-precipitation method. Detailed characterizations of NLDH such as its morphology, structure and particle size were performed by electron microscopy, X-ray diffraction（XRD） and dynamic light scattering technique（DLS）. The function of pure fluorouracil, fluorouracil and curcumin, fluorouracil loaded NLDH（LDH-Fu）, fluorouracil and curcumin co-loaded NLDH（LDH-Fu-Cur） on cell viability of 7721, LM3 and Hep G2 cell lines was measured by CCK-8 kit. Meanwhile, apoptosis kit was applied to detect apoptosis of7721 cells in the aforementioned groups. In order to explore the mechanism of apoptosis induced by fluorouracil and curcumin co-delivered NLDH, Western blot was used to detect the expression levels of apoptotic proteins such as Bcl-2, Caspase-3 and Caspase-9. Results NLDH, with a size of 400 nm, presented a good uniformity under electron microscope. XRD results indicated that fluorouracil and curcumin were horizontally inserted into the interlayer of NLDH. CCK8 results indicated that LDH-Fu-Cur more efficiently inhibited the growth of7721, LM3 and Hep G2 cell lines compared with pure fluorouracil, fluorouracil plus curcumin, and LDH-Fu（P〈0.05）. Similarly, LDH-Fu-Cur was more effective in promoting apoptosis than pure fluorouracil, fluorouracil plus curcumin, and LDH-Fu, as was evidenced by the apoptosis rate of LDH-Fu-Cur which was（87.0 ±4.7）% with the fluorouracil concentration of 35 μg/ml while the apoptosis rate of pure fluorouracil was only（23.0 ± 2.3）% with that concentration（P〈0.05）. Western blot results showed that NLDH could significantly down-regulate the Bcl-2 gene expression of 7721 cells and induce cell apoptosis by activating Caspase-3 and Caspase-9. Conclusions NLDH presents as a good carrier for anti-cancer drugs such as fluorouracil and curcumin and

关 键 词：层状双氢氧化物 肝细胞癌 氟尿嘧啶 姜黄素 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]