The cooperative effect of p53 and Rb in local nanotherapy in a rabbit VX2 model of hepatocellular carcinoma.
The cooperative effect of p53 and Rb in local nanotherapy in a rabbit VX2 model of hepatocellular carcinoma.
复制标题
p53和Rb在兔VX2肝细胞癌模型局部纳米治疗中的协同作用
DOI：
10.2147/ijn.s51353
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发表时间：
2013
影响因子：
8
通讯作者：
Li G
中科院分区：
文献类型：
--
作者：
Dong S;Tang Q;Long M;Guan J;Ye L;Li G
背景/目的一种局部纳米治疗（LNT）结合经动脉栓塞，纳米颗粒和p53基因治疗的治疗效果，以前已经提出。本研究旨在进一步改善LNT的不完全肿瘤根除和有限的生存提高，并阐明LNT的分子机制。方法以肿瘤靶向方式，将携带野生型p53和Rb的重组表达质粒在聚-L-赖氨酸修饰的羟基磷灰石纳米颗粒纳米复合物和Lipiodol®（Guerbet，Villepinte，France）乳剂中通过肝动脉共转移或分别转移到兔肝VX 2肿瘤中。随后的共表达p53和Rb蛋白内治疗的肿瘤进行了研究，通过Western印迹和原位分析，激光扫描共聚焦显微镜。通过肿瘤生长速度、凋亡率和坏死率、对阿霉素（ADM）、丝裂霉素C和氟尿嘧啶的敏感性、肿瘤组织微血管密度和动物存活时间来评价治疗效果。最后采用实时荧光定量聚合酶链反应和增强化学发光免疫印迹法检测治疗相关基因的表达变化。结果p53 + Rb LNT对兔的肝功能无明显影响，其抗肿瘤作用明显优于p53或Rb LNT，但对动物的生存率无明显影响。结论Rb与p53在聚赖氨酸修饰的羟基磷灰石纳米复合物介导的联合治疗中发挥协同作用，通过下调肿瘤细胞凋亡、坏死、生长、分化和多药耐药相关基因的表达增强抗肿瘤作用。联合p53和Rb的LNT是一种潜在的有效的抗肝癌治疗方法。
Background/aim A local nanotherapy (LNT) combining the therapeutic efficacy of trans-arterial embolization, nanoparticles, and p53 gene therapy has been previously presented. The study presented here aimed to further improve the incomplete tumor eradication and limited survival enhancement and to elucidate the molecular mechanism of the LNT. Methods In a tumor-targeting manner, recombinant expressing plasmids harboring wild-type p53 and Rb were either co-transferred or transferred separately to rabbit hepatic VX2 tumors in a poly-L-lysine-modified hydroxyapatite nanoparticle nanoplex and Lipiodol® (Guerbet, Villepinte, France) emulsion via the hepatic artery. Subsequent co-expression of p53 and Rb proteins within the treated tumors was investigated by Western blotting and in situ analysis by laser-scanning confocal microscopy. The therapeutic effect was evaluated by the tumor growth velocity, apoptosis and necrosis rates, their sensitivity to Adriamycin® (ADM), mitomycin C, and fluorouracil, the microvessel density of tumor tissue, and the survival time of animals. Eventually, real-time polymerase chain reaction and enhanced chemiluminescence Western blotting were used to investigate the expressive changes of important genes related to the therapy. Results The administration procedure proved safe for the rabbits’ liver function, the p53 plus Rb LNT showed significantly better antitumoral effect and lower expression of malignant genes than the p53 or Rb LNT, although no significant difference was observed in animal survival when the p53 plus Rb LNT was compared with the p53 LNT. Conclusion Rb works synergistically with p53 in combined therapy mediated by a poly-L-lysine-modified hydroxyapatite nanoparticle nanoplex to augment the antitumoral effect through the downregulated expression of important genes related to apoptosis, necrosis, growth, differentiation and multidrug resistance of tumor cells. LNT with p53 and Rb is potentially an effective antitumor therapy for hepatocellular carcinoma.