Development of New Strategy of Boron Neutron Capture Therapy to Cancer Combined with Tumor Specific Gene Delivery

我们对BNCT与肿瘤特异性基因治疗的增强效果进行了研究。(1)制备了一种聚乙二醇（PEG）结合脂质体，包裹了^<10>B化合物，用于递送系统。在热中子照射注入^<10> b -裸脂质体或^<10> b - peg -脂质体的小鼠后，与对照组相比，AsPC-1肿瘤的生长受到抑制。转铁蛋白-^<10> b - peg脂质体经BNCT检查，结肠癌肿瘤消失。体内热中子辐照注射转铁蛋白-^<10> b - peg脂质体和^<10> b - peg脂质体对肿瘤的抑制作用最大。(2)利用CR-39塑料径迹探测器进行中子俘获自放射照相（NCAR）。在立教大学，PSI， Jozef Stefan研究所或Sakley研究所，将样品附带的CR-39探测器暴露于热中子或冷中子中。利用聚乙二醇脂质体可以增加肿瘤组织中^<10 b> B原子的蓄积，从而增加其在血液中的潴留，使之更低，逃避res的吞噬作用。(3)研制了新型小型脉冲中子产生器。该中子发生器将应用于BNCT。(4)利用非病毒载体进行肿瘤基因治疗需要提高转染效率。我们制备了新型质粒DNA阳离子脂质体，称为四链复合体（Qplex），并评估了DNA的传递效率。Qplex由阳离子脂质体、pDNA、鱼精蛋白和转铁蛋白组成。脂质体M-（α-三甲基氨乙酰基）-二十二烷基-d -谷氨酰胺(TMAG) /二酰酰磷空间胆碱(DLPC) /二酰磷空间基乙醇胺（DOPE）（1:2:2）的脂质。在Xgal染色中，用Qplex转染lac Z基因的效率从常规阳离子脂质体的4%提高到52%。在血清存在的情况下，转染效率较高。(5)为了提高非病毒载体基因治疗的转染效率，在Lipoplex中对一种新型的融合原肽JTS-1进行了评价。使用JTS-1/LPD复合物的癌细胞荧光素酶活性最高。Xgal染色观察到转染效率高。LPD的平均直径为50 nm，与脂质体（约600nm）相比，它非常紧凑。使用JTS-1/LPD复合物的新型抑癌基因质粒tob为0.4μg / ml时，对AsPC-1细胞的肿瘤生长抑制率为50%。其抑制作用是诱导细胞凋亡。少

We performed the study of development of enhanced effects of BNCT with tumor specific gene therapy.(1) We prepare a polyethylene-glycol (PEG) binding liposome entrapped ^<10>B compound for the delivery system. After thermal neutron irradiation of mice injected with ^<10>B-bare liposome or ^<10>B-PEG-liposome, AsPC-1 tumour growth was suppressed relative to controls. Colon 26 tumor was disappeared by BNCT with the injection of Transferrin-^<10>B-PEG-Liposome. Injection of Transferrin-^<10>B-PEG-Liposome and ^<10>B-PEG-liposome caused the greatest tumour suppression with thermal neutron irradiation in vivo.(2) We performed neutron capture autoradiography(NCAR) using CR-39 plastic track detectors. The CR-39 detectors attached with samples were exposed to thermal neutrons or cold neutrons at Rikkyo University, PSI, Institute of Jozef Stefan or Sakley Institute. We can increase the accumulation of ^<10>B atoms in the tumor tissues by PEG-liposome, which increase the retension in the blood f … More low and escape the phagocytosis by RES.(3) We developed the novel small sized palsed neutron generator. This neutron generator will be applied to BNCT.(4) It is necessary to increase the transfectional efficiency when we use non-viral vector on cancer gene therapy. We have prepared new typed plasmid DNA-cationic lipoplexes, called quatenary complex (" Qplex "), and evaluated DNA delivery efficiency. Qplex is composed with cationic liposome, pDNA, protamine and transferrin. The lipid of liposome M-( α-trimethylammonioacetyl)-didodecyl-D-glutamatechloride (TMAG) / dilauroyl-phospatidylcholine (DLPC) / dioleoylphospatidyl-ethanolamine (DOPE) (1 : 2 : 2). The transfectional efficiency of lac Z gene is increased to 52% with Qplex from 4 % with conventional cationic lipoplexes in Xgal staining. The transfection efficiency is superior in the existence of serum.(5) In order to increase the transfection efficiency in non-viral vector gene therapy, a novel fusogenic peptide, JTS-1, was evaluated in Lipoplex. Luciferase activity in cancer cell lines was greatest in the using JTS-1/LPD complex. The high transfection effeciency was observed by Xgal staining. The mean diameter of LPD is 50 nm and it is so compact compared to lipoplex ( about 600nm ). Fifty percent tumor growth suppression of AsPC-1 cells was shown at the 0.4μg per ml of tob, "a novel tumor suppressor ", DNA plasmid using JTS-1/LPD complex. The suppressive effect was caused to induction of apoptosis. Less