Abstract: 【Abstract】〓Objective〓This work aims to investigate its effect on angiogenesis of hepatic colorectal metastasis,and to elucidate its molecular mechanism. Methods〓Recombinant expressing plasmids harboring wild type p53 and Rb were cotransferred or transferred separately to the mice hepatic VX2 tumor by the emulsion of Pll-nHAP nanoplex and lipodiol through the hepatic artery in a tumor target manner. Subsequent co-expressions of p53 and Rb protein within the treated tumors were investigated by western blotting and in situ analysis of confocal Laser scanning microscope. The therapeutic effect on angiogenesis was evaluated by the investigation of the microvessel density of tumor tissue. Eventually, the real-time RT-PCR and ECL western bolt were applied to investigate the expressive changes of important genes related to the above. Results〓(p53+Rb) LNT showed significantly better anti-angiogenesis effect and lower expression of related CD31, CD34, VEGF genes than the control group, although no significant difference was observed compared to p53 or Rb LNT for some genes. Conclusion〓Rb work synergistically with p53 in combined therapy mediated by Pll-nHAP nanoplex to augment the antitumoral effect through the down-regulated expression of important genes related to angiogenesis. Our study revealed that LNT with p53 and Rb is a potentially effective antitumor therapy for HCC.

Key words: Nanoparticles, Hepatic Metastasis, Gene Transfer Techniques, Rabbits

摘要： 【摘要】 目的 本研究旨在联合P53及Rb两种抗癌基因,通过纳米基因靶向治疗技术,探索其对肝转移癌灶血管新生的影响及其分子机制。方法〓以超液化碘油及Pll修饰的nHAP乳剂为载体,将含有野生型P53、Rb基因的重组表达质粒经肝动脉共同或者分别转运至兔VX2肝转移癌灶局部,采用蛋白印迹法及原位共焦激光显微镜确定P53及Rb共表达蛋白在转移灶的表达。然后病理观察肿瘤组织微血管密度(MVD)变化。最后应用Realtime RT-PCR级ECL蛋白杂交技术检测CD31、CD34及VEGF的表达差异。结果〓CD31标记的MVD显示:与其他组相比,nanoplex-p53/lipiodol及nanoplex-(p53+Rb)/lipiodol组肿瘤MVD均降低;CD34标记的MVD显示:与其他组相比,nanoplex- Rb/lipiodol及nanoplex-(p53+Rb)/lipiodol组MVD均降低。nanoplex- p53/lipiodol及nanoplex-(p53+Rb)/lipiodol组CD31 mRNA和蛋白表达降低;nanoplex- Rb/lipiodol及nanoplex-(p53+Rb)/lipiodol组CD34 mRNA和蛋白表达降低;nanoplex- p53/lipiodol、nanoplex- Rb/lipiodol及nanoplex-(p53+Rb)/lipiodol组VEGF mRNA和蛋白表达降低。结论〓以Pll-nHAP为载体的Rb基因协同P53基因纳米靶向治疗通过降低CD31、CD34及VEGF的表达减少新生血管生成,增强其抗肿瘤效应。P53基因联合Rb基因的纳米基因靶向治疗有可能成为抑制消化道癌肝转移灶血管新生的有效治疗方法。

关键词: 基因转染技术, 局部治疗, 肝转移癌, 兔